Granted Patent
B2
US 12,497,596 · App. 17/296,049 · Granted Dec 16, 2025
Hepatocyte expansion methods
C12N5/067A61K35/407A61P1/16C12N5/0671C12N2500/32C12N2500/38C12N2501/11C12N2501/117C12N2501/119C12N2501/12C12N2501/148C12N2501/15C12N2501/415C12N2501/727C12N2501/999C12N2503/04C12N2513/00C12N2533/90
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Abstract
This invention relates to culture methods and media for in vitro expansion of hepatocytes, particularly primary hepatocytes; hepatocyte cultures and organoids obtainable and obtained by said methods; and uses of said hepatocyte cultures and organoids.
Claims (42)
1 . A method for expanding hepatocytes in vitro to form a hepatocyte organoid, wherein the method comprises culturing isolated primary hepatocytes in a hepatocyte culture medium, wherein the hepatocyte culture medium comprises:
i) 50 to 500 ng/ml of a Fibroblast Growth Factor (FGF) selected from FGF7, FGF10, and a combination thereof;
ii) a Wnt agonist comprising an R-spondin selected from R-spondin 1, R-spondin 2, R-spondin 3, and R-spondin 4; and at least one glycogen synthase kinase 3 (GSK3) inhibitor;
iii) 5 to 100 ng/ml of epidermal growth factor (EGF);
iv) 5 to 100 ng/ml of hepatocyte growth factor (HGF); and
v) a transforming growth factor beta (TGF-β) inhibitor, comprising an inhibitor of activin receptor-like kinase ALK4, ALK5 and/or ALK7 signalling pathway;
wherein the hepatocyte organoid is formed, wherein the hepatocyte organoid does not express KRT7 and KRT19, and wherein the hepatocyte organoid expresses albumin.
2 . The method of claim 1 , wherein:
(a) the R-spondin is at a concentration of 5%-50% (vol/vol) within the hepatocyte culture medium;
(b) the GSK3 inhibitor is at a concentration of 1 to 10 μM, and wherein the GSK3 inhibitor is selected from: CHIR99021, SB216763, TWS119, 5-bromoindole, SB415286, BIO, AZD2858, AZD1080, AR-A014418, TDZD-8, LY2090314, BIO-acetoxime, 1-Azakenpaullone, and a combination thereof;
(c) the hepatocyte culture medium comprises FGF7 at a concentration of 50 to 400 ng/ml;
(d) the hepatocyte culture medium comprises FGF10 at a concentration of 50 to 400 ng/ml;
(e) the EGF is at a concentration of 10 ng/ml to 100 ng/ml;
(f) the HGF is at a concentration of 10 ng/ml to 100 ng/ml;
(g) wherein the TGF-β inhibitor is at a concentration of 1 to 10 μM, 0.5 to 5 μM, 1 to 4 μM, 1 to 3 μM, or about 2 μM; and/or
(h) the TGF-β inhibitor is an inhibitor of activin receptor-like kinase ALK4, ALK5 and/or ALK7 selected from: A83-01, SB-431542, SB-505124, EW-7197, LY-2157299, GW6604, and a combination thereof.
3 . The method of claim 1 , wherein:
(a) R-spondin 1 is at a concentration of about 15%;
(b) the GSK inhibitor is CHIR99021 at a concentration of about 3 μM; and/or
(c) the inhibitor of activin receptor-like kinase ALK4, ALK5 and/or ALK7 signaling pathway is A83-01 at a concentration of about 2 μM.
4 . The method of claim 1 , wherein the hepatocyte culture medium further comprises:
(a) nicotinamide at a concentration of 1 to 100 mM;
(b) gastrin at a concentration of 1 to 100 nM;
(c) transforming growth factor alpha (TGF-α) at a concentration of 1 to 50 ng/ml;
(d) a Rho-associated protein kinase (ROCK) inhibitor selected from: Y-27632, GSK429286A, Fasudil, Thiazovivin, Rho Kinase Inhibitor IV and a combination thereof, wherein the ROCK inhibitor is at a concentration of 1 to 50 μM;
(e) N-acetylcysteine, wherein the N-acetylcysteine is at a concentration of 0.5 to 5 mM; and/or
(f) a cell growth medium.
5 . The method of claim 1 , wherein the hepatocyte culture medium does not comprise a cAMP pathway activator.
6 . The method of claim 1 , wherein the hepatocytes are cultured in contact with a three dimensional extracellular matrix (ECM) in the presence of the hepatocyte culture medium.
7 . The method of claim 6 , wherein the three dimensional extracellular matrix is selected from: a synthetic ECM, a natural ECM, and a combination thereof.
8 . The method of claim 1 , wherein the hepatocytes comprise cells selected from: fetal hepatocytes, adult hepatocytes, and a combination thereof.
9 . The method of claim 1 , further comprising:
(a) expanding the hepatocyte organoids in culture for at least 10 passages; and/or
(b) expanding the hepatocyte organoids in culture for at least 3 months.
10 . The method of claim 1 , wherein:
(a) the hepatocyte organoids comprise a lumen; and/or
(b) the hepatocyte organoids secrete at least one of albumin, alpha-1 antitrypsin (A1AT), alpha-fetoglobulin (Afp), and a combination thereof.
11 . A method for expanding hepatocytes in vitro to form a hepatocyte organoid, wherein the method comprises culturing isolated primary hepatocytes in a hepatocyte culture medium,
wherein the hepatocyte culture medium comprises: about 100 ng/ml FGF7; about 100 ng/ml FGF10; about 20 ng/ml TGFa; about 50 ng/ml EGF; about 50 ng/ml HGF; about 2 μM A83-01; about 10 μM Y-27632; about 3 μM CHIR99021; about 10 mM nicotinamide; about 10 nM gastrin; about 1.25 mM N-acetylcysteine; and about 15% R-spondin conditioned medium;
wherein the hepatocyte organoid is formed,
wherein the hepatocyte organoid does not express KRT7 and KRT19, and
wherein the hepatocyte organoid expresses albumin.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 6, 2022
From: CLEVERS, JOHANNES CAROLUS; HU, HUILI
To: KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN
Reel/Frame 058566/0658 →
Priority Claims (1)
GB 1819224
· Nov 26, 2018
· national
Continuity (1)
References Cited (400)
US 5981483A
· Dennis et al.
· 1999
[cited by applicant]
US 6743626B2
· Baum et al.
· 2004
[cited by applicant]
US 8642339B2
· Sato et al.
· 2014
[cited by applicant]
US 8685726B2
· Schulz et al.
· 2014
[cited by applicant]
US 20030003088A1
· Tsao et al.
· 2003
[cited by applicant]
US 20030129751A1
· Grikscheit et al.
· 2003
[cited by applicant]
US 20030138951A1
· Yin
· 2003
[cited by applicant]
US 20040191902A1
· Hambor et al.
· 2004
[cited by applicant]
US 20040229355A1
· Chen et al.
· 2004
[cited by applicant]
US 20070010008A1
· Tseng et al.
· 2007
[cited by applicant]
US 20070036769A9
· Li et al.
· 2007
[cited by applicant]
US 20070122903A1
· Rezania et al.
· 2007
[cited by applicant]
US 20070128719A1
· Tseng et al.
· 2007
[cited by applicant]
US 20080182328A1
· Snyder et al.
· 2008
[cited by applicant]
US 20080233088A1
· Guha et al.
· 2008
[cited by applicant]
US 20080242594A1
· McKay et al.
· 2008
[cited by applicant]
US 20090275067A1
· Taniguchi et al.
· 2009
[cited by applicant]
US 20100100396A1
· Daven et al.
· 2010
[cited by applicant]
US 20110008893A1
· Sugimura et al.
· 2011
[cited by applicant]
US 20110191868A1
· Gupta et al.
· 2011
[cited by applicant]
US 20120207744A1
· Mendlein et al.
· 2012
[cited by applicant]
US 20130052729A1
· Pourquie et al.
· 2013
[cited by applicant]
US 20130089562A1
· French et al.
· 2013
[cited by applicant]
US 20140044713A1
· Lau et al.
· 2014
[cited by applicant]
US 20140243227A1
· Clevers et al.
· 2014
[cited by applicant]
US 20140302491A1
· Nadauld et al.
· 2014
[cited by applicant]
US 20150140013A1
· Ramaswamy
· 2015
[cited by applicant]
US 20150276719A2
· Beekman et al.
· 2015
[cited by applicant]
US 20160002595A1
· Keller et al.
· 2016
[cited by applicant]
US 20180072995A1
· Sato et al.
· 2018
[cited by applicant]
US 20180187191A1
· Zeng
· 2018
[cited by applicant]
US 20180258400A1
· Ng et al.
· 2018
[cited by applicant]
US 20190100728A1
· Sato et al.
· 2019
[cited by applicant]
US 20200377860A1
· Freedman et al.
· 2020
[cited by applicant]
US 20210047618A1
· Clevers et al.
· 2021
[cited by applicant]
US 20220340879A1
· De Lau et al.
· 2022
[cited by applicant]
US 20240076624A1
· Beumer et al.
· 2024
[cited by applicant]
AU 2012265814A2
· 2012
[cited by applicant]
CN 101420964A
· 2009
[cited by applicant]
CN 103180436A
· 2013
[cited by applicant]
CN 103237888A
· 2013
[cited by applicant]
CN 104508121A
· 2015
[cited by applicant]
EP 0953633A1
· 1999
[cited by applicant]
EP 2412800A1
· 2012
[cited by applicant]
EP 2420566A1
· 2012
[cited by applicant]
EP 2772534A1
· 2014
[cited by applicant]
EP 2138571B1
· 2017
[cited by applicant]
EP 3318627A1
· 2018
[cited by applicant]
EP 3505620B1
· 2023
[cited by applicant]
JP 2012000097A
· 2012
[cited by applicant]
WO 2004050827A2
· 2004
[cited by applicant]
WO 2004087896A3
· 2004
[cited by applicant]
WO 2005040391A1
· 2005
[cited by applicant]
WO 2005120547A1
· 2005
[cited by applicant]
WO 2007141657A2
· 2007
[cited by applicant]
WO 2007127454A3
· 2008
[cited by applicant]
WO 2008046649A1
· 2008
[cited by applicant]
WO 2008101215A1
· 2008
[cited by applicant]
WO 2009022907A2
· 2009
[cited by applicant]
WO 2009024595A2
· 2009
[cited by applicant]
WO 2010011352A2
· 2010
[cited by applicant]
WO 2010015938A2
· 2010
[cited by applicant]
WO 2010016766A2
· 2010
[cited by applicant]
WO 2010090513A1
· 2010
[cited by applicant]
WO 2010090513A2
· 2010
[cited by applicant]
WO 2010119819A1
· 2010
[cited by applicant]
WO 2010129294A3
· 2011
[cited by applicant]
WO 2011098402A1
· 2011
[cited by applicant]
WO 2011043591A2
· 2011
[cited by applicant]
WO 2012014076A2
· 2012
[cited by applicant]
WO 2012025725A1
· 2012
[cited by applicant]
WO 2012044992A2
· 2012
[cited by applicant]
WO 2012068251A2
· 2012
[cited by applicant]
WO 2012014076A3
· 2012
[cited by applicant]
WO 2012087965A3
· 2012
[cited by applicant]
WO 2012140274A2
· 2012
[cited by applicant]
WO 2012168930A2
· 2012
[cited by applicant]
WO 2012140274A9
· 2013
[cited by applicant]
WO 2013054112A1
· 2013
[cited by applicant]
WO 2013061608A1
· 2013
[cited by applicant]
WO 2013093812A2
· 2013
[cited by applicant]
WO 2013074681A9
· 2013
[cited by applicant]
WO 2014015777A1
· 2014
[cited by applicant]
WO 2014066649A1
· 2014
[cited by applicant]
WO 2014124527A1
· 2014
[cited by applicant]
WO 2014127170A1
· 2014
[cited by applicant]
WO 2014127219A1
· 2014
[cited by applicant]
WO 2014145389
· 2014
[cited by applicant]
WO 2014159356A1
· 2014
[cited by applicant]
WO 2014170411A1
· 2014
[cited by applicant]
WO 2015040142A1
· 2015
[cited by applicant]
WO 2015173425A1
· 2015
[cited by applicant]
WO 2015179393A1
· 2015
[cited by applicant]
WO 2015200901A1
· 2015
[cited by applicant]
WO 2016016894A1
· 2016
[cited by applicant]
WO 2016056999A1
· 2016
[cited by applicant]
WO 2016083612A1
· 2016
[cited by applicant]
WO 2016083613A2
· 2016
[cited by applicant]
WO 2016094457A1
· 2016
[cited by applicant]
WO 2017048193A1
· 2017
[cited by applicant]
WO 2017120543A1
· 2017
[cited by applicant]
WO 2017149025A1
· 2017
[cited by applicant]
WO 2017205511A1
· 2017
[cited by applicant]
WO 2017220586A1
· 2017
[cited by applicant]
WO 2018035138A1
· 2018
[cited by applicant]
WO 2018052953A1
· 2018
[cited by applicant]
WO WO2018036119A1
· 2018
[cited by examiner]
WO 2018102202A1
· 2018
[cited by applicant]
WO 2019122388A1
· 2019
[cited by applicant]
WO 2019228516A1
· 2019
[cited by applicant]
WO 2020205755A1
· 2020
[cited by applicant]
WO 2020234250A1
· 2020
[cited by applicant]
Park et al. Differential activities and mechanisms of the four R-spondinsin potentiating Wnt/b-catenin signaling. J Biol Chem .Jun. 22, 2018;293(25):9759-9769. Epub May 11, 2018. (Year: 2018).
[cited by examiner]
Drost et al., “Organoid culture systems for prostate epithelial and cancer tissue”, Nat. Protoc, Jan. 2016 (Jan. 21, 2016), 11(2): 347-358 (25 pages).
[cited by applicant]
Drost et al., “Sequential cancer mutations in cultured human intestinal stem cells,” Nature, vol. 521, May 7, 2015 (23 pages).
[cited by applicant]
Duncan et al., “The ploidy-conveyor of mature hepatocytes as a source of genetic variation,” Nature. Oct. 7, 2010; 467(7316): 707-710 (14 pages).
[cited by applicant]
Eccles, “The epidermal growth factor receptor/Erb-B/HER family in normal and malignant breast biology,” International Journal of Developmental Biology, University of the Basque Country Pres., 55(7-9): 685-696 (Jan. 1, 2…
[cited by applicant]
Eden et al., “Gorilla: a tool for discovery and visualization of enriched GO terms in ranked gene lists,” BMC Bioinformatics 2009, 10:48.
[cited by applicant]
Egerod et al., “A Major Lineage of Enteroendocrine Cells Coexpress CCK, Secretin, Gip, GLP-1, PYY, and Neurotensin but Not Somatostatin Endocrinology,” Dec. 2012, 153(12):5782-5795.
[cited by applicant]
Engelhardt et al., “Detection of α-foetoprotein in mouse liver differentiated hepatocytes before their progression through S phase,” 1976 Nature 263: 146-148.
[cited by applicant]
English translation of Official Action issued in Russian Application No. RU2018145539A, dated Mar. 19, 2021 (8 pages).
[cited by applicant]
English translation of Search Report issued in Russian Application No. RU2018145539A, dated Mar. 19, 2021 (2 pages).
[cited by applicant]
Evarts et al., “A precursor—product relationship exists between oval cells and hepatocytes in rat liver,” Carcinogenesis 8(11): 1737-1740, 1987 (4 pages).
[cited by applicant]
Fan et al. “Cholangiocarcinomas can originate from hepatocytes in mice,” J Clin Invest, Aug. 2012, 122(8): 2911-2915 (5 pages).
[cited by applicant]
Farin et al., “Basic and Translational-Alimentary Tract,” (2012) Gastroenterology, 143:1518-1529.
[cited by applicant]
Farin et al., “Visualization of a short-range Wnt gradient in the intestinal stem-cell niche,” Nature, 340 Feb. 18, 2016 (15 pages).
[cited by applicant]
Finnberg et al., “Application of 3D tumoroid systems to define immune and cytotoxic therapeutic responses based on tumoroid and tissue slice culture molecular signatures”, Oncotarget, Sep. 15, 2017, 8(40): 66747-66757 (…
[cited by applicant]
Finnberg et al., Abstract 3990: “Use of 3D tumoroid systems to define immune and cytotoxic therapeutic responses based on tumoroid and tissue slice culture molecular signatures : Cancer Research”, Proceedings of the Ame…
[cited by applicant]
Font-Burgada et al., “Hybrid Periportal Hepatocytes Regenerate the Injured Liver without Giving Rise to Cancer,” 2015 Cell 162: 766-779 (15 pages).
[cited by applicant]
Fuchs, Ota, “Inhibition of TGF-B Signaling for the Treatment of Tumor Metastatis and Fibrotic Diseases”, Bentham Science Publishers Ltd.; (2011); 15 pages.
[cited by applicant]
Furuyama et al., “Continuous cell supply from a Sox9-expressing progenitor zone in adult liver, exocrine pancreas and intestine,” 2011 Nat Genet 43: 34-41 (11 pages).
[cited by applicant]
Gao et al., “3D Spheroid/Organoid Models of Lung Cancer to Study Lung Cancer Pathogenesis and Testing of New Therapeutics”, Journal of Thoracic Oncology, vol. 12, S1544, 1 page (Jun. 21, 2017).
[cited by applicant]
Gerbal-Chaloin et al., “The WNT/b-Catenin Pathway Is a Transcriptional Regulator of CYP2E1, CYP1A2, and Aryl Hydrocarbon Receptor GeneExpression in Primary Human Hepatocytes,” (2014) Molecular Pharmacology 86:624-634.
[cited by applicant]
Ghosh et al., “Activity Assay of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors in Triple-Negative Breast Cancer Cells Using Peptide-Conjugated Magnetic Beads,” Assay and Drug Development Technologies, 11(1…
[cited by applicant]
Greene et al., “Partial Hepatectomy in the Mouse: Technique and Perioperative Management,” 2003 J Invest Surg 16: 99-102 (4 pages).
[cited by applicant]
Grompe, M., “Liver Stem Cells, Where Art Thou?,” Cell Stem Cell 15: 257-258, 2014 (2 pages).
[cited by applicant]
Grompe,M. “Fah Knockout Animals as Models for Therapeutic Liver Repopulation,” Hereditary Tyrosinemia, Advances in Experimental Medicine and Biology, 2017, 959: 215-230 (16 pages).
[cited by applicant]
Grun et al., “Single-cell messenger RNA sequencing reveals rare intestinal cell types,” Nature 525: 251-255 (Sep. 10, 2015).
[cited by applicant]
Gui et al., “Heregulin protects mesenchymal stem cells from serum deprivation and hypoxia-induced apoptosis,” Mol Cell Biochem, 305:171-178 (2007).
[cited by applicant]
Gunawardene et al., “Classification and functions of enteroendocrine cells of the lower gastrointestinal tract, ” Int. J. Exp. Path. (2011), 92: 219-231.
[cited by applicant]
Harada et al., “Rapid formation of hepatic organoid in collagen sponge by rat small hepatocytes and hepatic nonparenchymal cells,” Journal of Hepatology (2003) 39: 716-723.
[cited by applicant]
Haramis et al., “De novo crypt formation and juvenile polyposis on BMP inhibition in mouse intestine.” Science Mar. 12, 2004, 303(5664): 1684-1686.
[cited by applicant]
Hashimshony et al., “CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification,” Cell Reports 2: 666-673, Sep. 27, 2012.
[cited by applicant]
Hashimshony et al., “CEL-Seq2: sensitive highly-multiplexed single-cell RNA-Seq,” Genome Biol 17: 77, 2016 (7 pages).
[cited by applicant]
Hayashi et al., “Establishment and characterization of a parietal endoderm-like cell line derived from Engelbreth-Holm-Swarm tumor (EHSPEL), a possible resource for an engineered basement membrane matrix,” Science Direc…
[cited by applicant]
Heuberger et al., “Shp2/MAPK signaling controls goblet/paneth cell fate decisions in the intestine,” PNAS 111(9): 3472-3477 (Mar. 4, 2014).
[cited by applicant]
Hirshhaeuser et al., “Efficacy of catumaxomab in tumor spheroid killing is mediated by its trifunctional mode of action,” Cancer Immunol Immunother, Jul. 2010, 59: 1675-1684 (10 pages).
[cited by applicant]
Hofer and Drenckhahn, “Cytoskeletal markers allowing discrimination between brush cells and other epithelial cells of the gut including enteroendocrine cells,” Cell Biol (1996) 105:405-412.
[cited by applicant]
Homback-Klonisch et al., Adult stem cells and their trans-differentiation potential-perspectives and therapeutic applications. J Mol Med (Berl). 2008; 86(12):1301-1314 (26 pages).
[cited by applicant]
Jou et al., “Pluripotent Stem Cells Induced from Mouse Somatic Cells by Small-Molecule Compounds,” Science, 341 (6146):651-654, science.org/doi/10.1126/science. 1239278; 8 pages (Jul. 18, 2013).
[cited by applicant]
Howitt et al., “Tuft cells, taste-chemosensory cells, orchestrate parasite type 2 immunity in the gut,” Science. Mar. 18, 2016; 351(6279): 1329-1333.
[cited by applicant]
Hsieh et al., “Truncated Mammalian Notch1 Activates CBF1/RBPJk-Repressed Genes by a Mechanism Resembling That of Esptein-Barr Virus EBNA2,” Molecular and Cellular Biology, (Mar. 1996). pp. 952-959.
[cited by applicant]
Hu et al., “Direct Conversion of Normal and Alzheimer's Disease Human Fibroblasts into Neuronal Cells by Small Molecules,” Cell Stem Cell 17: 204-212, Aug. 6, 2015 (36 pages).
[cited by applicant]
Huang et al., “Direct Reprogramming of Human Fibroblasts to Functional and Expandable Hepatocytes,” Cell Stem Cell 14: 370-384, Mar. 6, 2014 (15 pages).
[cited by applicant]
Huang et al., “Induction of functional hepatocyte-like cells from mouse fibroblasts by defined factors,” Nature Jul. 21, 2011, 475: 386-389 (7 pages).
[cited by applicant]
Huch & Clevers, “Sox9 marks adult organ progenitors,” Nature Genetics, Jan. 11, 2011, 43(1): 9-10 (3 pages).
[cited by applicant]
Huch et al., In vitro expansion of single Lgr51 liver stem cells induced by Wnt-driven regeneration 2013 Nature 494: 247-250 (7 pages).
[cited by applicant]
Huch et al., Long-Term Culture of Genome-Stable Bipotent Stem Cells from Adult Human Liver, (2015) Cell 160: 299-312 (24 pages).
[cited by applicant]
Huch et al., “Urokinase-Type Plasminogen Activator Receptor Transcriptionally Controlled Adenoviruses Eradicate Pancreatic Tumors and Liver Metastasis in Mouse Models,” NeoPlasia, 11(6): 518-528 (Jun. 2009).
[cited by applicant]
Huschtscha et al., “Normal human mammary epithelial cells proliferate rapidly in the pressure of elevated levels of the tumor suppressors p53 and p21WAF1/CIP1,” Journal of Cell Science, 122: 2989-2995 (2009).
[cited by applicant]
Hynds et al., “The Relevance of Human Stem Cell-Derived Organoid Models for Epithelial Translational Medicine,” Europe PMC Funders Group Stem Cells (Mar. 2013); 11 pages.
[cited by applicant]
Garashi et al., “Characterization of Recombinant Human Fibroblast Growth Factor (FGF)-10 Reveals Functional Similarities with Keratinocyte Growth Factor (FGF-7),” The Journal of Biological Chemistry, (May 1998) 273(21):…
[cited by applicant]
International Preliminary Report on Patentability of International Application No. PCT/EP2019/082618, dated Jun. 10, 2021 (13 pages).
[cited by applicant]
U.S. Appl. No. 16/078,354, filed Aug. 21, 2018, Johannes Carolus Clevers.
[cited by applicant]
U.S. Appl. No. 16/310,933, filed Dec. 18, 2018, Johannes Carolus Clevers.
[cited by applicant]
U.S. Appl. No. 17/915,771, filed Sep. 29, 2022, Joep Beumer
[cited by applicant]
Besser et al., “Modifying interleukin—2 concentrations during culture improves function of T cells for adoptive immunotherapy,” Cytotherapy (2009) 11(2): 206-217 (12 pages).
[cited by applicant]
Beumer et al., “High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells”, Cell, May 13, 2020, 181(6): 1291-1306 (40 pages).
[cited by applicant]
Chang-Graham et al., “Human Intestinal Enteroids With Inducible Neurogenin-3 Expression as a Novel Model of Gut Hormone Secretion”, Cellular and Molecular Gastroenterology, Sep. 10, 2019, 8(2): 209-229 (21 pages).
[cited by applicant]
Dwyer et al., “A three-dimensional co-culture system to investigate macrophage-dependent tumor cell invasion,” Journal of Biological Methods, vol. 3(3), e49, 10 pages (2016).
[cited by applicant]
Gehart et al., “Identification of Enteroendocrine Regulators by Real-Time Single-Cell Differentiation Mapping,” Cell 176: 1158-1173, Feb. 21, 2019 (33 pages).
[cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/EP2021/058527, dated Aug. 25, 2021 (23 pages).
[cited by applicant]
Neal et al., “Organoid Modeling of the Tumor Immune Microenvironment,” Cell. Dec. 13, 2018; 175(7):1972-1988.e16. doi: 10.1016/j.cell.2018.11.021 (34 pages).
[cited by applicant]
Sinagoga et al., “Deriving functional human enteroendocrine cells from pluripotent stem cells,” Development (2018) 145, dev165795, doi:10.1242/dev.165795 (11 pages).
[cited by applicant]
Santhanam et al., “Upregulated Pathways and Products of Tryptophan Metabolism is Associated with the Neoplastic Transition in the Colon Epithelium,” Gastroenterology 2016, 150, 4, suppl 1, p. S-492, Abstract Su1195(1 pa…
[cited by applicant]
Casey et al., “Theory of cell fate,” WIREs Syst Biol Med, 2020; 12:e1471 (11 pages).
[cited by applicant]
Supek et al., “REVIGO Summarizes and Visualizes Long Lists of Gene Ontology Terms,” PLoS ONE 6(7): 2011; e21800, 9 pages.
[cited by applicant]
Swenson, E., “Direct Conversion of Mouse Fibroblasts to Hepatocyte-Like Cells Using Forced Expression of Endodermal Transcription Factors,” 2012 Hepatology 55(1): 316-318 (9 pages).
[cited by applicant]
Takeda et al., “Inter-conversion between intestinal stem cell populations in distinct niches,” Science. Dec. 9, 2011; 334(6061): 1420-1424.
[cited by applicant]
Tanimizu et al., “Notch signaling controls hepatoblast differentiation by altering the expression of liver-enriched transcription factors,” Journal of Cell Science, 2014, 117(15): 3165-3174.
[cited by applicant]
Tanimizu et al., “Sry HMG Box Protein 9-positive (Sox9+) Epithelial Cell Adhesion Molecule-negative (EpCAM-) Biphenotypic Cells Derived from Hepatocytes Are Involved in Mouse Liver Regeneration,” J Biol Chem, 289(11): 7…
[cited by applicant]
Tarlow et al., “Bipotential Adult Liver Progenitors Are Derived from Chronically Injured Mature Hepatocytes,” Nov. 6, 2014, Cell Stem Cell 15: 605-618 (15 pages).
[cited by applicant]
Terry et al., “Impaired enteroendocrine development in intestinal-specific Islet1 mouse mutants causes impaired glucose homeostasis,” Am J Physiol Gastrointest Liver Physiol 307: G979-G991, Sep. 11, 2014.
[cited by applicant]
Tetteh et al., “Replacement of Lost Lgr5-Positive Stem Cells through Plasticity of Their Enterocyte-Lineage Daughters,” Cell Stem Cell 18: 203-213, Feb. 4, 2016.
[cited by applicant]
“The Wnt Family of Secreted Proteins”, R&D Systems' 2004 Catalog, 7 pages (Jan. 1, 2004).
[cited by applicant]
Thenappan et al., New Therapeutics Targeting Colon Cancer Stem Cells, Curr Colorectal Cancer Rep, vol. 5, No. 4, 2009 (12 pages).
[cited by applicant]
Tian et al., “Integrin-specific hydrogels as adaptable tumor organoids for malignant Band T cells”, Biomaterials, Sep. 11, 2015, 73: 110-119 (21 pages).
[cited by applicant]
Tisato et al., “Upregulation of SOCS-1 by Nutlin-3 in acute myeloid leukemia cells but not in primary normal cells,” Clinics (2014) pp. 68-74.
[cited by applicant]
Tojo et al., “The ALK-5 inhibitor A-83-01 inhibits Smad signaling and epithelial-to-mesenchymal transition bytransforming growth factor-beta,” Cancer Sci, 2005, 96(11): 791-800.
[cited by applicant]
Trierweiler et al., “The transcription factor c-JUN/AP-1 promotesHBV-related liver tumorigenesis in mice,” Cell Death and Differentiation 23: 576-582 (2016).
[cited by applicant]
Tsai et al., “LGR4 and LGR5 Function Redundantly During Human Endoderm Differentiation”, Cellular and Molecular Gastroenterology and Hepatology, 2016, 2: 648-662.
[cited by applicant]
Upton et al., “De novo synthesis of T cells from mPB CD34+ cells cultured in a 3-dimensional thymic organoid,” Blood 102(11): 279a, Nov. 16, 2003 (2 pages).
[cited by applicant]
Van Amerongen et al., “Developmental Stage and Time Dictate the Fate of Wnt/β-Catenin-Responsive Stem Cells in the Mammary Gland,” Sep. 7, 2012, Cell Stem Cell 11: 387-400 (14 pages).
[cited by applicant]
Van de Wetering et al., “Prospective Derivation of a Living Organoid Biobank of Colorectal Cancer Patients,” Cell, 161, May 7, 2015, 161: 933-945 (14 pages).
[cited by applicant]
Van Es et al., “DII1 marks early secretory progenitors in gut crypts that can revert to stem cells upon tissue damage,” Nat Cell Biol. Oct. 2012; 14(10): 1099-1104.
[cited by applicant]
Van Es et al., “Notch/γ-Secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells,” vol. 435; 5 pages, Jun. 2005.
[cited by applicant]
Vaughan et al., “Lineage-negative Progenitors Mobilize to Regenerate Lung Epithelium after Major Injury,” HHS Public Access Nature (Jan. 29, 2015); 25 pages.
[cited by applicant]
Verbeke et al., “Humanization of the mouse mammary gland by replacement of the luminal layer with genetically engineered preneoplastic human cells,” Breast Cancer Research (2014) 16: 504, 20 pages.
[cited by applicant]
Verkhovskaya et al. Deystvie alkoksizameshchyonnykh glitserina na morfofunktsionalnye svoystva perevivaemoy kultury (Effect of Alkoxy-substituted of Glycerol on Morphofunctional Properties of a Passaged Culture), Kriobi…
[cited by applicant]
Verma et al., “Sustained Telomere Length in Hepatocytes and Cholangiocytes with Increasing Age in Normal Liver,” 2012, Hepatology 56:1510-1520 (11 pages).
[cited by applicant]
Vickaryous et al., “Human cell type diversity, evolution, development, and classification with special reference to cells derived from the neural crest,”: Biol Rev Camb Philos Soc., 81(3):425-55, Aug. 2006 (31 pages).
[cited by applicant]
Vincan et al., “Frizzled-7 dictates three-dimensional organization of colorectal cancer cell carcinoids”, Oncogene 2007, 26: 2340-2352.
[cited by applicant]
Voronkov et al., “Wnt/beta-Catenin Signaling and Small Molecule Inhibitors,” Current Pharmaceutical Design, 19: 634-664 (2013).
[cited by applicant]
Wang et al., “Self-renewing diploid Axin21 cells fuel homeostatic renewal of the liver,” Aug. 13, 2015, Nature, 524: 180-185 (18 pages).
[cited by applicant]
Williams et al., “The Role of the Wnt Family of Secreted Proteins in Rat Oval Stem” Cell-Based Liver Regeneration Wnt1 Drives Differentiation, American Journal of Pathology, Jun. 2010, 176(6): 2732-2742.
[cited by applicant]
Wouters et al., “Evolution of distinct EGF domains with specific functions,” Protein Science (2005) 14: 1091-1103.
[cited by applicant]
Wu et al., “FGF19 Regulates Cell Proliferation, Glucose and Bile Acid Metabolism vai FGFR4-Dependent and Independent Pathways,” PLOS One, Mar. 2011, 6(3): e17868, pp. 1-11.
[cited by applicant]
Yan et al., “The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations,” PNAS 109 (2): 466-471 (Jan. 10, 2012).
[cited by applicant]
Yang et al., “Differentiation of Human Induced-Pluripotent Stem Cells into Smooth-Muscle Cells: Two Novel Protocols,” PLoS One, 2016; 11(1):e0147155 (11 pages).
[cited by applicant]
Yang et al., “In vitro trans-differentiation of adult hepatic stem cells into pancreatic endocrine hormoneproducing cells,” PNAS, Jun. 11, 2002, 99(12): 8078-8083.
[cited by applicant]
Yang et al., “Beta-catenin signaling in murine liver zonation and regeneration: A Wnt-Wnt situation!” (2014) Hepatology 60(3): 964-976.
[cited by applicant]
Yanger et al., “Adult Hepatocytes Are Generated by Self-Duplication Rather than Stem Cell Differentiation,” Sep. 4, 2014, Cell Stem Cell 15: 340-349 (10 pages).
[cited by applicant]
Yanger et al., “Robust cellular reprogramming occurs spontaneously during liver regeneration,” 2013, Genes Dev 27: 719-724 (8 pages).
[cited by applicant]
Yimlamai et al., “Hippo Pathway Activity Influences Liver Cell Fate,” Jun. 5, 2014, Cell, 157(6): 1324-1338 (23 pages).
[cited by applicant]
Yin et al., “Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny,” Nat Methods. Jan. 2014; 11(1): 106-112.
[cited by applicant]
Yokoyama et al., “Regeneration of Mouse Liver after Partial Hepatectomy,” 1953 Cancer Res 13: 80-85 (7 pages).
[cited by applicant]
Yoshimura et al., “Vascular endothelial cells and smooth muscle cells mediate carbachol-induced hepatocyte proliferation via muscarinic receptors and IP3/PKC signaling cascades,” Cell Biol Int. Apr. 2009; 33(4): 516-23.
[cited by applicant]
Yu et al., “Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility,” HHS Public Access Science (Jul. 11, 2014); 345(6193): 216-220.
[cited by applicant]
Zaret et al., “Genetic programming of liver and pancreas progenitors: lessons for stem-cell differentiation,” Nature Reviews, May 2008, 9: 329-340.
[cited by applicant]
Zauli et al., “MDM2 Antagonist Nutlin-3 Suppresses the Proliferation and Differentiation of Human Pre-Osteoclasts Through a p53-Dependent Pathway,” Journal of Bone and Mineral Research vol. 22, No. 10; (2007) 10 pages.
[cited by applicant]
Zhu et al., “Chemical Strategies for Stem Cell Biology and Regenerative Medicine”, The Annual Review of Biomedical Engineering Apr. 20, 2011, 13(1): 73-90.
[cited by applicant]
Zhu et al., “Mouse liver repopulation with hepatocytes generated from human fibroblasts,” Apr. 3, 2014, Nature 508 (7494): 93-97 (34 pages).
[cited by applicant]
Zilberberg et al., “A Rapid and Sensitive Bioassay to Measure Bone Morphogenetic Protein Activity,” BMC Cell Biology BioMed Central (Sep. 19, 2007), vol. 8, Article 41, 10 pages.
[cited by applicant]
Zimmerman, “Lung organoid culture,” Differentiation; Research in Biological Diversity, 36(1): 86-109 (1987).
[cited by applicant]
Zong et al., “Notch signaling controls liver development by regulating biliary differentiation,” Development 136: 1727-1739 (2009).
[cited by applicant]
Zuo et al., “P63 + Krt5 + distal airway stem cells are essential for lung regeneration,” Nature (2014) 517:616-620, 17 pages.
[cited by applicant]
Basak et al., “Induced quiescence of Lgr5+ stem cells in intestinal organoids enables differentiation of hormone-producing enteroendocrine cells,” Cell Stem Cell.; 20 (2), 177-190.e4 (2017).
[cited by applicant]
Beumer et al., “Enteroendocrine cells switch hormone expression along the crypt-to-villus BMP signalling gradient,” Nature Cell Biology. 20 (8), pp. 909-916 (2018).
[cited by applicant]
Blume et al., “Temperature corrected transepithelial electrical resistance (TEER) measurement to quantify rapid changes in paracellular permeability,” Die Pharmazie; 65 (1), pp. 19-24 (2010).
[cited by applicant]
Boj et al., “Forskolin-induced swelling in intestinal organoids: An in vitro assay for assessing drug response in cystic fibrosis patients,” Journal of Visualized Experiments, 2017 (120), pp. 1-12 (2017).
[cited by applicant]
De Lau et al., “The R-spondin protein family,” Genome Biology. 13:242, 2012 (10 pages).
[cited by applicant]
Dinter et al., “Pharmacologic strategies for assaying BMP signaling function,” (2019) Methods Mol Biol; 1891:221-233 (13 pages).
[cited by applicant]
Drakos et al., “Inhibition of p53-Murine Double Minute 2 Interaction by Nutlin-3A Stabilizes p53 and Induces Cell Cycle Arrest and Apoptosis in Hodgkin Lymphoma,” Clin Cancer Res 2007; 13 (11), Jun. 1, 2007, pp. 3380-33…
[cited by applicant]
Grimaldi et al., A Cell Model Suitable for a High-Throughput Screening of Inhibitors of the Wnt/β-catenin pathway, Frontiers in Pharmacology, vol. 9, Article 1160, Oct. 2018 (3 pages).
[cited by applicant]
Gupta et al., “Scaffolding kidney organoids on silk,” Tissue Eng Regen Med. May 2019 ; 13(5): 812-822 (20 pages).
[cited by applicant]
Hosic et al., “Cholinergic Activation of Primary Human Derived Intestinal Epithelium Does Not Ameliorate TNF-a Induced Injury,” Cellular and Molecular Bioengineering, vol. 13, No. 5, Oct. 2020 (2020) pp. 487-505 (19 pag…
[cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/EP2022/069354, dated Nov. 1, 2022, by Examiner (14 pages).
[cited by applicant]
Kucab et al., “Selection of TP53-mutated human TP53 knock-in (Hupki) mouse embryo fibroblasts using the MDM2 inhibitor Nutlin-3a,” Mutagenesis, vol. 29 No. 1 pp. 79-96, Abstracts of the 36th Annual Meeting of the United…
[cited by applicant]
Leonard et al., “Screening of Budesonide Nanoformulations for Treatment of Inflammatory Bowel Disease in an Inflamed 3D Cell-Culture Model,” Altex, 29(3):275-285, 2012 (11 pages).
[cited by applicant]
Martínez-Maqueda, et al., “HT29 Cell Line. in The Impact of Food Bio-Actives on Gut Health: In Vitro and Ex Vivo Models,” Verhoeckx, K. et al. (eds), Cham 20 (CH): Springer, 113-124 (2015).
[cited by applicant]
Miao, Y. et al., “Next-generation surrogate Wnts support organoid growth and deconvolute Frizzled pleiotropy in vivo.” Cell Stem Cell. 27 (5), 840-851 (2020).
[cited by applicant]
Miyachi et al., “Restoration of p53 Pathway by Nutlin-3 Induces Cell Cycle Arrest and Apoptosis in Human Rhabdomyosarcoma Cells,” Clin Cancer Res, 15(12), Jun. 15, 2009, pp. 4077-4084 (8 pages).
[cited by applicant]
Noben et al., “Human intestinal epithelium in a dish: Current models for research into gastrointestinal pathophysiology,” United European Gastroenterology Journal 2017, vol. 5(8) 1073-1081 (9 pages).
[cited by applicant]
Nusse et al., Wnt/β-Catenin Signaling, Disease, and Emerging Therapeutic Modalities, Jun. 1, 2017, Cell 169 (6):985-999.
[cited by applicant]
Romier et al., “New cell-based HTRF® assays for the exploration of Wnt signalling pathway” Cisbio Bioassays, 2015 (1 page).
[cited by applicant]
Sanchez-Duffhues, “Bone morphogenetic protein receptors: Structure, function and targeting by selective small molecule kinase inhibitors,” (2020) Bone 138:115472 (13 pages).
[cited by applicant]
Schmohl et al., “Characterization of immunologically active drugs in a novel organotypic co-culture model of the human gut and whole blood,” International Immunopharmacology 14 (2012) pp. 722-728 (7 pages).
[cited by applicant]
Schutgens et al., “Tubuloids derived from human adult kidney and urine for personalized disease modeling,” Nature Biotechnology 37: 303-313, (2019).
[cited by applicant]
Singh et al., “Review on EGFR Inhibitors: Critical Updates,” (2016) Mini-Reviews in Medicinal Chemistry 16:1134-1166.
[cited by applicant]
Snyder et al., “Materials and Microenvironments for Engineering the Intestinal Epithelium,” Ann Biomed Eng., vol. 48, No. 7, Feb. 4, 2020, pp. 1916-1940.
[cited by applicant]
Srinivasan et al., “TEER measurement techniques for in vitro barrier model systems.” Journal of Laboratory 5 Automation. 20 (2), 107-126 (2015).
[cited by applicant]
Tran and Zheng, “Modulating the wnt signaling pathway with small molecules,” (2017) Protein Science 26:650-661 (12 pages).
[cited by applicant]
Trenker and Jura, “Receptor tyrosine kinase activation: From the ligand perspective,” Current Opinion in Cell Biology, 2020, 63:174-185 (12 pages).
[cited by applicant]
Vadstrup et al., “Ex Vivo Assay of Intestinal Mucosal Biopsies in Crohn's Disease: Reflects Inflammation and Drug Effects,” PLoS One, May 2016 (18 pages).
[cited by applicant]
Van der Vaart et al., “Modelling of primary ciliary dyskinesia using patient-derived airway organoids,” EMBO reports; 2021; 22: e52058 (16 pages).
[cited by applicant]
Van Es et al., “Wnt signalling induces maturation of Paneth cells in intestinal crypts,” Nature Cell Biology. 7(4), 381-386 (2005).
[cited by applicant]
Vanuytsel et al., “Major Signaling Pathways in Intestinal Stem Cells,” Biochim Biophys Acta. Feb. 2013; 1830(2): 2410-2426 (42 pages).
[cited by applicant]
Villalonga-Planells et al., “Activation of p53 by Nutlin-3a Induces Apoptosis and Cellular Senescence in Human Glioblastoma Multiforme,” PLoS One, vol. 6, Issue 4, Apr. 2011, e18588 (12 pages).
[cited by applicant]
Zhao Y, et al. Cancer Research, 101st Annual Meeting of the American Association for Cancer Research, AACR 2010, Washington, DC, United States, Apr. 17, 2010-21, (Apr. 15, 2010) vol. 70, No. 8, Supp. Suppl. 1. Abstract …
[cited by applicant]
Kohno et al., “Effects of hyaluronidase on doxorubicin penetration into squamous carcinoma multicellular tumor spheroids and its cell lethality,” J Cancer Res Clin Oncol. 1994; 120(5):293-7 (6 pages).
[cited by applicant]
Koo et al., “Controlled gene expression in primary Lgr5 organoid cultures,” Nat Methods. 2012 9(1): 81-84 (4 pages).
[cited by applicant]
Kramer et al., “Small-Molecule Inhibitors of GSK-3: Structural Insights and Their Application to Alzheimer's Disease Models,” International Journal of Alzheimer's Disease, vol. 2012, Article ID 381029, 32 pages.
[cited by applicant]
Kross et al., “Co-culture of Head and Neck Squamous Cell Carcinoma Spheroids with Autologous Monocytes Predicts Prognosis,” Scand J Immunol. 2008; 67(4):392-9 (8 pages).
[cited by applicant]
Laban et al., “Sorafenib sensitizes head and neck squamous cell carcinoma cells to ionizing radiation,” Radiotherpy and Oncology, 2013, 109(2): 286-292 (7 pages).
[cited by applicant]
Lau et al., “The R-spondin protein family,” Genome Biol. 2012; 13(3):242, 2012 (10 pages).
[cited by applicant]
Leemans et al., “The molecular landscape of head and neck cancer,” May 2018, Nat. Rev. Cancer, vol. 18(5): 269-282 (14 pages).
[cited by applicant]
Lengauer et al., “Genetic instability in colorectal cancers,” Nature. Apr. 10, 1997; 386: 623-627 (5 pages).
[cited by applicant]
Li and Durbin, “Fast and accurate long-read alignment with Burrows-Wheeler transform,” Bioinformatics. 2010, 26(5): 589-595 (7 pages).
[cited by applicant]
Li et al., “Conformational equilibria and intrinsic affinities define integrin activation,” (2017) EMBO 36:629-645 (17 pages).
[cited by applicant]
Luque et al., “Activated Conformations of Very Late Activation Integrins Detected by a Group of Antibodies (HUTS) Specific for a Novel RegulatoryRegion (355-425) of the Common β1 Chain,” (1996) J. Bio Chem. 271(19): 110…
[cited by applicant]
Machiels et al., “Activity and safety of afatinib in a window preoperative EORTC study in patients with squamous cell carcinoma of the head and neck (SCCHN),” Annals of Oncology 29: 985-991, 2018 (7 pages).
[cited by applicant]
Machiels et al., “Afatinib versus methotrexate as second-line treatment in patients with recurrent or metastatic squamous-cell carcinoma of the head and neck progressing on or after platinum-based therapy (LUX-Head & Ne…
[cited by applicant]
Martin et al., “Modeling antibody hypervariable loops: A combined algorithm,” Proc. Natl. Acad. Sci. USA, 86: 9268-9272, Dec. 1989 (5 pages).
[cited by applicant]
Maushagen et al., “Effects of paclitaxel on permanent head and neck squamous cell carcinoma cell lines and Identification of anti-apoptotic caspase 9b,” J Cancer Res Clin Oncol. 2016; 142(6):1261-71.
[cited by applicant]
Mayer et al., “A Phase Ib Study of Alpelisib (BYL719), a PI3Kα-Specific Inhibitor, with Letrozole in ER+/HER2− Metastatic Breast Cancer,” Metastatic Breast Cancer, Clin Cancer Res. Jan. 1, 2017, 23(1): 26-34 (10 pages).
[cited by applicant]
Meng et al., “Characterization of integrin engagement during defined human embryonic stem cell culture,” (2010) The FASEB Journal, 24(4): 1056-1065 (17 pages).
[cited by applicant]
Méry et al., “Preclinical models in HNSCC: A comprehensive review,” Oral Oncol. 2017, 65: 51-56 (6 pages).
[cited by applicant]
Monnet et al., “Selection of IgG variants with increased FcRn binding using random and directed mutagenesis: impact on effector functions,” Frontiers in Immunology, vol. 6, Article 39, Feb. 2015 (15 pages).
[cited by applicant]
Munster et al., “Abstract A46: Inhibition of PIK3CA with BYL719 canovercome resistance to cetuximab in squamous cellcarcinoma of the head and neck (SCCHN),” Mol Cancer Ther. Jul. 2015; vol. 14, Issue 7 Supplement (4 pag…
[cited by applicant]
Pokharel et al., “Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response,” Nature Communications, (2019) 10:1482 (17 pages).
[cited by applicant]
Polychronopoulos et al., Structural Basis for the Synthesis of Indirubins as Potent and Selective Inhibitors of Glycogen Synthase Kinase-3 and Cyclin-Dependent Kinases, (2004) J Med Chem 47: 935-946 (12 pages).
[cited by applicant]
Pyeon et al., “Production of infectious human papillomavirus independently of viral replication and epithelial cell differentiation,” Proc Natl Acad Sci. 2005; 102(26):9311-9316 (6 pages).
[cited by applicant]
Reed, “Plantinum-DNA adduct, nucleotide excision repair and platinum based anti-cancer chemotherapy,” Cancer Treat. Rev. 1998. p. 331-44 (14 pages).
[cited by applicant]
Sachs et al., “A Living Biobank of Breast Cancer Organoids Captures Disease Heterogenity,” Cell 172: 373-386 (Jan. 11, 2018).
[cited by applicant]
Sachs et al., “Long-term expanding human airway organoids for disease modeling,” The EMBO Journal (2019) 38: e100300 (20 pages).
[cited by applicant]
Sayers and Elliott, Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature J Virol. 2016; 90(22):10379-10389 (11 pages).
[cited by applicant]
Schrader et al., “Kallikrein-related peptidase 6 regulates epithelial-to-mesenchymal transition and serves as prognostic biomarker for head and neck squamous cell carcinoma patients,” Mol Cancer. 2015; 14:107 (14 pages).
[cited by applicant]
Shah et al., “Metabolic Imaging of Head and Neck Cancer Organoids,” PLoS ONE, 2017, 12(1):e0170415 (17 pages).
[cited by applicant]
Shaner et al., “A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum,” Nat Methods. 2013 10(5): 407-414 (8 pages).
[cited by applicant]
Shattil et al., “The final steps of integrin activation: the end game,” (Apr. 2010) Nature reviews, 11: 288-300 (13 pages).
[cited by applicant]
Shimizu et al., “Identification of a novel therapeutic target for head and neck squamous cell carcinomas: A role for the heurotensin-neurotensin receptor 1 oncogenic signaling pathway,” Int J Cancer. 2008; 123(8):1816-1…
[cited by applicant]
Simmini S et al., “Transformation of intestinal stem cells into gastric stem cells on loss of transcription factor Cdx2,” Nat Commun. 2014; 5:5728 (10 pages).
[cited by applicant]
Smith et al., “Animal models for the study of squamous cell carcinoma of the upperaerodigestive tract: A historical perspective with review of their utility and limitations. Part A. Chemically-induced de novo cancer, sy…
[cited by applicant]
Smits et al., “Immortalized N/TERT keratinocytes as an alternative cell source in 3D human epidermal models,” Sci Rep. 2017; 7(1):11838 (14 pages).
[cited by applicant]
Soulières D et al., “Buparlisib and paclitaxel in patients with platinum-pretreated recurrent or metastatic squamous cell carcinoma of the head and neck (BERIL-1): a randomised, double-blind, placebo-controlled phase 2 …
[cited by applicant]
Squier et al., “Biology of Oral Mucosa and Esophagus,” J Natl Cancer Inst Monogr. 2001; 29:7-15 (9 pages).
[cited by applicant]
Su et al., “Relating conformation to function in integrin α5β1,” (2016) PNAS 113(27): E3872-E3881 (10 pages).
[cited by applicant]
Sun et al., “Integrin activation by talin, kindlin and mechanical forces,” Nature Cell Biology, Jan. 2019, 21: 25-31 (7 pages).
[cited by applicant]
Tanaka et al., “Head and neck cancer organoids established by modification of the CTOS method can be used to predict in vivo drug sensitivity,” Oral Oncology. 2018, 87: 49-57 (9 pages).
[cited by applicant]
The Cancer Genome Atlas Network, “Comprehensive genomic characterization of head and neck squamous cell carcinomas,” Nature. Jan. 29, 2015; 517: 576-582 (7 pages).
[cited by applicant]
Tsuchida et al., “Classification of ‘activation’ antibodies against integrin BI chain,” (1997) FEBS Letters 416: 212-21 (5 pages).
[cited by applicant]
Valyi-Nagy T et al., “Herpes Simplex Virus 1 Infection Promotes the Growth of a Subpopulation of Tumor Cells in Three-Dimensional Uveal Melanoma Cultures,” J Virol. Oct. 2018; 92(19): e00700-18 (12 pages).
[cited by applicant]
Van Jaarsveld RH et al., “Difference Makers: Chromosomal Instability versus Aneuploidy in Cancer,” Trends in cancer. 2(10): 561-571 (11 pages).
[cited by applicant]
Vassilev et al., In Vivo Activation of the p53 Pathway by Small-Molecule Antagonist of MDM2, Science. 2004; 303 (5659):844-848 (7 pages).
[cited by applicant]
Vlachogiannis et al., “Patient-derived organoids model treatment response of metastatic gastrointestinal cancers,” Science. Feb. 23, 2018; 359(6378): 920-926 (17 pages).
[cited by applicant]
Voskoglou-Nomikos et al., “Clinical Predictive Value of the in Vitro Cell Line, Human Xenograft, and Mouse Allograft Preclinical,” Cancer Models1Clin Cancer Res. 2003, 9(11):4227-39 (14 pages).
[cited by applicant]
Yan et al., “Expression profile analysis of head and neck squamous cell carcinomas using data from The Cancer Genome Atlas,” Mol Med Rep. 2016; 13(5):4259-4265 (7 pages).
[cited by applicant]
Yang et al., A Small Molecule Agonist of an Integrin, αLβ2*,S, Yang et al.(2006) Journal of biological chemistry, 281(49): 37904-37912 (18 pages).
[cited by applicant]
Yu et al., β1-Integrin Orients Epithelial Polarity via Rac1 and Laminin, (Feb. 2005) Molecular Biology of the cell, 16: 433-445 (13 pages).
[cited by applicant]
International Search Report and Written Opinion for International Application No. PCT/EP2015/060815, dated Jul. 28, 2015 (15 pages).
[cited by applicant]
International Search Report and Written Opinion for International Application No. PCT/EP2015/077988, dated Apr. 20, 2016 (13 pages).
[cited by applicant]
International Search Report and Written Opinion for International Application No. PCT/EP2015/077990, mailed Jul. 6, 2016 (17 pages).
[cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/EP2017/054797, dated May 31, 2017 (9 pages).
[cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/EP2017/065101, dated Oct. 6, 2017 (13 pages).
[cited by applicant]
International Search Report and Written Opinion of International Application No. PCT/EP2019/082618, dated Feb. 11, 2020 (15 pages).
[cited by applicant]
Janssen, and Depoortere, “Nutrient sensing in the gut: new roads to therapeutics? Trends in endocrinology and metabolism,” 24: 92-100 (2013).
[cited by applicant]
Jeong et al., “Neuregulin-1 induces cancer stem cell characteristics in breast cancer cell lines,” Oncology Reports (2014), 32: 1218-1224.
[cited by applicant]
Kamiya et al., “Oncostatin M and hepatocyte growth factor induce hepatic maturation via distinct signaling pathways,” FEBS Letters 2001, 492: 90-94 (5 pages).
[cited by applicant]
Kan et al., “MPp53-mediated growth suppression in response to Nutlin-3 in cyclin D1 transformed,” Cancer Research (Nov. 2007) 9 pages.
[cited by applicant]
Katsuda et al., “Conversion of Terminally Committed Hepatocytes to Culturable Bipotent Progenitor Cells with Regenerative Capacity,” Cell Stem Cell 20: 41-55, Jan. 5, 2017 (16 pages).
[cited by applicant]
Ke et al. “Down-regulation of Wnt signaling could promote bone marrow derived mesenchymal stem cells to differentiate into hepatocytes,” BBRC, Jan. 2, 2008, 36: 342-348 (7 pages).
[cited by applicant]
Kemp et al., “The Roles of Wnt Signaling in Early Mouse Development and Embryonic Stem Cells”, Functional Development and Embryology, 2007, pp. 1-13.
[cited by applicant]
Khetani et al., “Microscale culture of human liver cells for drug development,” Nat Biotechnol 26(1): 120-126 (20 pages).
[cited by applicant]
Kim et al., “Mitogenic Influence of Human R-Spondin1 on the Intestinal Epithelium” Science, Aug. 19, 2005, 309: 1256-1259.
[cited by applicant]
Kirikoshi et al., “WNT10A and WNT6, Clustered in Human Chromosome 2q35 Region with Head-to-Tail Manner, Are Strongly Coexpressed in SW480 Cells,” Biochemical and Biophysical Research Communications (2001), 283: 798-805.
[cited by applicant]
Kogata et al., “Neuregulin 3 and Erbb Signalling Networks in Embryonic Mammary Gland Development,” J Mammary Gland Biology Neoplasia (2013) 18: 149-154.
[cited by applicant]
Koo et al., “Stem Cells Marked by the R-Spondin Receptor LGR5,” Gastroenterology 2014; 147:289-302.
[cited by applicant]
Morin et al., “Activation of β-Catenin-Tcf-Signaling in Colon Cancer by Mutations in β-Catenin or APC,” Science, 275: 1787-1790, Mar. 21, 1997.
[cited by applicant]
Korinek et al., “Constitutive Transcriptional Activation by a β-Catenin-Tcf Complex in APC -/- Colon Carcinoma,” (1997) Science 275:1784-1787.
[cited by applicant]
Kuball et al., “Facilitating matched pairing and expression of TCR chains doi:10.1182/blood introduced into human T cells,” Blood, 109(6): 2331-2338, 8 pages (2007).
[cited by applicant]
Latorre et al., “Enteroendocrine Cells: A Review of Their Role In Brain-Gut Communication,” Neurogastroenterol Motil. May 2016; 28(5): 620-630.
[cited by applicant]
Lee et al., “Lung stem cell differentiation in mice directed by endothelial cells via a BMP4-NFATc1-Thrombospondin-1 axis,” Cell. Jan. 30, 2014; 156(3): 440-455.
[cited by applicant]
Lee et al., “Neuregulin Autocrine Signaling Promotes Self-Renewal of Breast Tumor-Initiating Cells by Triggering HER2/HER3 Activation,” Tumor and Stem Cell Biology Cancer Research 74(1):341-52 (2014).
[cited by applicant]
Lemaigre et al., “Mechanisms of Liver Development: Concepts for Understanding Liver Disorders and Design of Novel Therapies”, Gastroenterology Jul. 1, 2009, (137)(1): 62-79.
[cited by applicant]
Levy et al., “Long-term culture and expansion of primary human hepatocytes,” 2015 Nat Biotechnol 33: 1264-1271 (10 pages).
[cited by applicant]
Li et al., “Adult Mouse Liver Contains Two Distinct Populations of Cholangiocytes,” Stem Cell Reports, 9: 478-489, Aug. 8, 2017 (12 pages).
[cited by applicant]
Li et al., “Hepatoblast-Like Progenitor Cells Derived From Embryonic Stem Cells Can Repopulate Livers of Mice,” Gastroenterology 139: 2158-2169, e2158 2010 (20 pages).
[cited by applicant]
Li et al., “Isolation and Culture of Adult Mouse Hepatocytes,” Methods Mol Biol, 633: 185-196, 2010 (12 pages).
[cited by applicant]
Liang et al., “Genetic and Epigenetic Variations in iPSCs: Potential Causes and Implications for Application,” Cell Stem Cell 13: 149-159, Aug. 1, 2013 (11 pages).
[cited by applicant]
Lin et al., “Distributed hepatocytes expressing telomerase repopulate the liver in homeostasis and injury,” Nature 556: 244-248, Apr. 12, 2018 (7 pages).
[cited by applicant]
Little et al., “Engineering Biomaterials for Synthetic Neural Stem Cell Microenvironments,” (2008) Chem. Rev 108, 1787-1796.
[cited by applicant]
Liu et al., “Osteopontin Promotes Hepatic Progenitor Cell Expansion and Tumorigenicity via Activation of β-Catenin in Mice,” 2015 Stem Cells 33: 3569-3580 (12 pages).
[cited by applicant]
Love et al., “Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2,” Genome Biology, 15: 550, 2014 (21 pages).
[cited by applicant]
Lund et al., “Genetic and epigenetic stability of human pluripotent stem cells,” 2012, Nat Rev Genet 13: 732-744 (14 pages).
[cited by applicant]
Lyzikov A.N. et al., Stvolovye kletki v regenerativnoy meditsine: dostizheniya i perspektivy (Stem Cells in Regenerative Medicine: Achievements and Prospects), Problemy zdorovya i ekologii, 2015.
[cited by applicant]
Macchiarini et al., “Clinical Transplantation of a Tissue-Engineered Airway,” The Lancet (Nov. 19, 2008) 8 pages.
[cited by applicant]
Malato et al., “Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration,” Dec. 2011, J Clin Invest, 121(12): 4850-4860 (11 pages).
[cited by applicant]
Malorni et al., “The antioxidant N-acetyl-cysteine protects cultured epithelial cells from menadione-induced cytopathology”, Chemico-Biological Interactions 1995, 96: 113-123.
[cited by applicant]
Manandhar et al., “Glucagon-like Peptide-1 (GLP-1) Analogs: Recent Advances, New Possibilities, and Therapeutic Implications,” J. Med. Chem. 2015, 58: 1020-1037.
[cited by applicant]
Marquardt et al., “Functional and genetic deconstruction of the cellular origin in liver cancer,” Nov. 2015, J Nat Rev Cancer 15: 653-667 (15 pages).
[cited by applicant]
Martin-Belmonte et al., “Cell-Polarity Dynamics Controls the Mechanism of Lumen Formation in Epithelial Morphogenesis”, Current Biology, 2008, 18: 507-513.
[cited by applicant]
McEwen et al., “Regulation of the Fibroblast Growth Factor Receptor 3 Promoter and Introm I Enhancer by Sp1 Family Transcription Factors,” The Journal of Biological Chemistry, 273(9): 5349-5357 (Feb. 27, 1998).
[cited by applicant]
MediLexicon Dictionary, http://www.medilexicon.com/medicaldictionary.php?t=63274 , “Organoid”, 2006, p. 1.
[cited by applicant]
Michalopoulos, G., “Liver Regeneration after Partial Hepatectomy,” Jan. 2010, Am J Pathol 176(1): 2-13 (13 pages).
[cited by applicant]
Mitaka, Toshihiro, “Reconstruction of hepatic organoid by hepatic stem cells”, J. Hepatobiliary Pancreat Sug, Jan. 1, 2002, 9(6): 697-703.
[cited by applicant]
Miyajima et al., “Stem/Progenitor Cells in Liver Development, Homeostasis, Regeneration, and Reprogramming,” May 1, 2014, Cell Stem Cell 14: 561-574 (14 pages).
[cited by applicant]
Munoz et al., “The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent ‘+4’ cell markers,” The EMBO Journal (2012) 31: 3079-3091.
[cited by applicant]
Murry et al. “Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development,” Cell, 22;132(4):661-80, Feb. 2008 (20 pages).
[cited by applicant]
Ahmed et al., “Extracellular Matrix Regulation of Stem Cell Behavior,” Curr Stem Cell Rep (2016) 2:197-206 (10 bages).
[cited by applicant]
Al-Lazikani et al., “Standard Conformations for the Canonical Structures of Immunoglobulins,” J. Mol. Biol. (1997) 273: 927-948 (22 pages).
[cited by applicant]
Andersson et al., “Pharmacokinetics of Cisplatin and Its Monohydated Coplex in Humans,” J Pharm Sci. Aug. 1996; 85(8): 824 827 (4 pages).
[cited by applicant]
Argiris et al., “Head and neck cancer,” Lancet. May 17, 2008; 371(9625): 1695-1709 (32 pages).
[cited by applicant]
Atsushi et al., A comparison of in vitro platinum-DNA adduct formation between carboplatin and cisplatin, Int J Biochem. 1994; 26(8): 1009-1016 (8 pages).
[cited by applicant]
Barretina et al., “The CancerCell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity,” Nature, 2012; 483(7391): 603-607 (8 pages).
[cited by applicant]
Bhosale et al., “Chromosomal Alterations and Gene Expression Changes Associated with the Progression of eukoplakia to Advanced Gingivobuccal Cancer,” Transl Oncol. 2017; 10(3):396-409 (14 pages).
[cited by applicant]
Bigorgne et al., “TTC7A mutations disrupt intestinal epithelial apicobasal polarity,” J Clin Invest. 2014; 124(1):328-337 (11 pages).
[cited by applicant]
Boj et al., “Organoid Models of Human and Mouse Ductal Pancreatic Cancer,” Cell. Jan. 15, 2015; 160(1-2): 324-338 (28 pages).
[cited by applicant]
Bossi et al., “Prognostic and predictive value of EGFR in head and neck squamous cell carcinoma,” Oncotarget. 2016; 7(45): 74362-74379 (18 pages).
[cited by applicant]
Braakhuis et al., “The Potential of the Nude Mouse Xenograft Model for the Study of Head and Neck Cancer,” Arch Otorhinolaryngol. 1984, 239(1): 69-79 (11 pages).
[cited by applicant]
Byron et al.“Anti-integrin monoclonal antibodies,” J Cell Sci. Nov. 15, 2009; 122(Pt 22): 4009-4011 (6 pages).
[cited by applicant]
Cai Y et al., “Dysregulations in the PI3K pathway and targeted therapies for head and neck squamous cell carcinoma,” Oncotarget. 2017; 8 (13): 2203-22217 (15 pages).
[cited by applicant]
Calderwood et al., “Integrin activation,” Journal of Cell Science 117(5): 657-666, 2004 (10 pages).
[cited by applicant]