IP Library › Granted Patent US 12,247,188
Granted Patent B2
US 12,247,188 · App. 17/077,447 · Granted Mar 11, 2025

Cell culture chamber with improved cell-contacting surfaces

Inventors: Nuala Trainor (Kingston, CA); Eytan Abraham (Walkersville, MD); Timothy Smith (Kingston, CA); Matthew Hewitt (Walkersville, MD); Yaling Shi (Walkersville, MD); Kelly Purpura (Kingston, CA); Chase McRobie (Kingston, CA)
Assignees: OCTANE BIOTECH INC.; LONZA WALKERSVILLE, INC.
C12M23/04C12M23/20C12M23/24C12M23/42C12M27/00C12M29/24C12M41/12C12M41/32C12M41/34C12M41/48C12N5/0068G01N35/1009C12N2533/52
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,247,188
App. No.
17/077,447
Granted
Mar 11, 2025
Kind
B2
Abstract

The present disclosure provides cell culture chambers for use in automated cell engineering systems, and in particular, cell culture chambers that include improved cell-contacting surfaces. Improved cell-contacting surfaces can include a surface coating that promotes cell growth, adherence, differentiation, maintenance of phenotype, and/or improves transduction; a cell-contacting surface comprising a non-porous, gas-permeable material; as well as other modifications to the cell-contacting surfaces. Cassettes comprising the cell culture chambers are also provided.

Claims (36)

1. A cassette for use in an automated cell engineering system as a removable and replaceable element, comprising:

a. a high temperature chamber configured for carrying out activation, transduction and/or expansion of a cell culture at a temperature of between 35° C. to 40° C., the high temperature chamber including a cell culture chamber;

b. a low temperature chamber configured for storage of a reagent at a temperature of between 4° C. to 8° C.;

c. a thermal barrier adjacent the high temperature chamber and the low temperature chamber and separating the high temperature chamber from the low temperature chamber within the cassette; and

d. one or more fluidics pathways connected to the cell culture chamber, wherein the fluidics pathways provide recirculation, removal of waste and homogenous gas exchange and distribution of nutrients to the cell culture chamber without disturbing cells within the cell culture chamber;

wherein the cell culture chamber is a flat and non-flexible chamber, having a low chamber height and a cell-contacting surface, and the cell culture chamber is maintained in a substantially planar orientation in the cassette, and

wherein at least a portion of the cell-contacting surface comprises a non-porous, gas-permeable material, and the cell-contacting surface further includes multiple structures cut or made into the cell-contacting surface to add surface area to increase cell adhesion, wherein the multiple structures are sandwiched between sheets of the non-porous, gas-permeable material.

2. The cassette of claim 1 , wherein the cell culture chamber further comprises at least one of:

i. a distal port configured to allow for the removal of air bubbles from the cell culture chamber and/or as a recirculation port;

ii. a medial port configured to function as a recirculation inlet port; and

iii. a proximal port configured to function as a drain port for cell removal.

3. The cassette of claim 1 , wherein the cell-contacting surface comprises a plurality of separate sections, each section comprising the non-porous, gas-permeable material.

4. The cassette of claim 1 , wherein at least about 50% of the cell-contacting surface comprises the non-porous, gas-permeable material.

5. The cassette of claim 1 , wherein the entire cell-contacting surface comprises the non-porous, gas-permeable material.

6. The cassette of claim 1 , wherein the non-porous, gas-permeable material comprises silicone, flouroethylenepolypropylene (FEP), or ethyl vinyl olefin (EVO).

7. The cassette of claim 1 , having a chamber height of about 0.5 cm to about 4 cm.

8. The cassette of claim 1 , having a volume of about 50 ml to about 200 ml.

9. The cassette of claim 1 , wherein the cassette does not include a centrifugation chamber.

10. The cassette of claim 1 , wherein the cell-contacting surface further comprises a surface coating on the cell-contacting surface selected from the group consisting of:

i. a surface coating that activates a cell;

ii. a surface coating that modulates a biological pathway in a cell;

iii. a surface coating that enhances growth of a cell;

iv. a surface coating that improves adhesion of a cell;

V. a surface coating that inhibits a cell;

vi. a surface coating that responds to media conditions; and

vii. a surface coating that has controlled solubility.

11. The cassette of claim 10 , wherein the surface coating comprises an adhesion molecule.

12. The cassette of claim 11 , wherein the adhesion molecule is fibronectin or a modified fibronectin.

13. The cassette of claim 1 , wherein the cassette is pre-filled with culture media, activation reagent, and optionally a vector.

14. The cassette of claim 1 , further comprising one or more of a pH sensor, a glucose sensor, an oxygen sensor, a lactate sensor, a cell counting module, a carbon dioxide sensor, and/or an optical density sensor.

15. The cassette of claim 1 , further comprising one or more sampling ports and/or injection ports.

16. The cassette of claim 1 , further comprising an access port for connecting the cassette to an external device.

17. The cassette of claim 16 , wherein the external device includes an electroporation unit or an additional media source.

18. The cassette of claim 1 , wherein one or more of the fluidic pathways comprise a silicone-based tubing component that allows oxygenation through the tubing component.

19. The cassette of claim 1 , wherein a portion of the cell-contacting surface further comprises a surface treatment.

20. The cassette of claim 1 , wherein the low temperature chamber further includes a bag or other holder for the reagent.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 13, 2026
From: LONZA WALKERSVILLE, INC.
To: OCTANE BIOTECH INC.
Reel/Frame 075379/0734 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 3, 2020
From: ABRAHAM, EYTAN; HEWITT, MATTHEW; SHI, YALING
To: LONZA WALKERSVILLE, INC.
Reel/Frame 054532/0315 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 3, 2020
From: TRAINOR, NUALA; SMITH, TIMOTHY; PURPURA, KELLY; MCROBIE, CHASE
To: OCTANE BIOTECH INC.
Reel/Frame 054532/0363 →
Continuity (2)
Provisional Application 62925392 · Oct 24, 2019
Related Publication 20210123008A1 · Apr 29, 2021
References Cited (157)
US 4642120A · Nevo et al. · 1987 [cited by applicant]
US 4939151A · Bacehowski et al. · 1990 [cited by applicant]
US 5041138A · Vacanti et al. · 1991 [cited by applicant]
US 5081036A · Familletti · 1992 [cited by applicant]
US 5240854A · Berry et al. · 1993 [cited by applicant]
US 5246699A · Debre et al. · 1993 [cited by applicant]
US 5424209A · Kearney · 1995 [cited by applicant]
US 5478479A · Herrig · 1995 [cited by applicant]
US 5549134A · Browne et al. · 1996 [cited by applicant]
US 5688687A · Palsson et al. · 1997 [cited by applicant]
US 5728581A · Schwartz et al. · 1998 [cited by applicant]
US 5786207A · Katz et al. · 1998 [cited by applicant]
US 5792603A · Dunkelman et al. · 1998 [cited by applicant]
US 5827729A · Naughton et al. · 1998 [cited by applicant]
US 5842477A · Naughton et al. · 1998 [cited by applicant]
US 5846828A · Peterson et al. · 1998 [cited by applicant]
US 5882929A · Fofonoff et al. · 1999 [cited by applicant]
US 5891455A · Sittinger et al. · 1999 [cited by applicant]
US 5902741A · Purchio et al. · 1999 [cited by applicant]
US 5906934A · Grande et al. · 1999 [cited by applicant]
US 5922604A · Stapleton et al. · 1999 [cited by applicant]
US 5928936A · Ingram · 1999 [cited by applicant]
US 5935847A · Smith · 1999 [cited by examiner]
US 5985653A · Armstrong et al. · 1999 [cited by applicant]
US 5989913A · Anderson et al. · 1999 [cited by applicant]
US 5994129A · Armstrong et al. · 1999 [cited by applicant]
US 6048721A · Armstrong et al. · 2000 [cited by applicant]
US 6048722A · Farb et al. · 2000 [cited by applicant]
US 6060306A · Flatt et al. · 2000 [cited by applicant]
US 6096532A · Armstrong et al. · 2000 [cited by applicant]
US 6121042A · Peterson et al. · 2000 [cited by applicant]
US 6123655A · Fell · 2000 [cited by applicant]
US 6197575B1 · Griffith et al. · 2001 [cited by applicant]
US 6214574B1 · Kopf · 2001 [cited by applicant]
US 6228635B1 · Armstrong et al. · 2001 [cited by applicant]
US 6238908B1 · Armstrong et al. · 2001 [cited by applicant]
US 6297046B1 · Smith et al. · 2001 [cited by applicant]
US 6323146B1 · Pugh et al. · 2001 [cited by applicant]
US 6402941B1 · Lucido et al. · 2002 [cited by applicant]
US 7348175B2 · Vilendrer et al. · 2008 [cited by applicant]
US 7560274B1 · Fuller · 2009 [cited by examiner]
US 7906323B2 · Cannon et al. · 2011 [cited by applicant]
US 9629877B2 · Cooper et al. · 2017 [cited by applicant]
US 10131876B2 · Kaiser et al. · 2018 [cited by applicant]
US 10253316B2 · Masquelier et al. · 2019 [cited by applicant]
US 10273300B2 · Bedoya et al. · 2019 [cited by applicant]
US 11208626B2 · Mason et al. · 2021 [cited by applicant]
US 20010021529A1 · Takagi · 2001 [cited by applicant]
US 20010043918A1 · Masini et al. · 2001 [cited by applicant]
US 20020009797A1 · Wolf et al. · 2002 [cited by applicant]
US 20020009803A1 · Gabor Vajta · 2002 [cited by applicant]
US 20020025547A1 · Rao · 2002 [cited by applicant]
US 20020037580A1 · Schoeb · 2002 [cited by applicant]
US 20020146816A1 · Vellinger et al. · 2002 [cited by applicant]
US 20020155487A1 · Greenberger et al. · 2002 [cited by applicant]
US 20020179525A1 · Shaffer et al. · 2002 [cited by applicant]
US 20030008388A1 · Barbera-Guillem · 2003 [cited by examiner]
US 20030032071A1 · Wang et al. · 2003 [cited by applicant]
US 20030040104A1 · Barbera-Guillem · 2003 [cited by applicant]
US 20030054335A1 · Taya et al. · 2003 [cited by applicant]
US 20030159946A1 · Eden et al. · 2003 [cited by applicant]
US 20030215935A1 · Coon · 2003 [cited by applicant]
US 20040048364A1 · Trosch · 2004 [cited by applicant]
US 20050064465A1 · Dettloff et al. · 2005 [cited by applicant]
US 20050106717A1 · Wilson · 2005 [cited by examiner]
US 20050130297A1 · Sarem et al. · 2005 [cited by applicant]
US 20050186671A1 · Cannon et al. · 2005 [cited by applicant]
US 20080227176A1 · Wilson · 2008 [cited by examiner]
US 20090148941A1 · Florez · 2009 [cited by examiner]
US 20100055774A1 · Wilson · 2010 [cited by examiner]
US 20110198286A1 · Niazi · 2011 [cited by examiner]
US 20130059339A1 · Karerangabo · 2013 [cited by examiner]
US 20130084030A1 · Staheli · 2013 [cited by examiner]
US 20140120608A1 · Carter · 2014 [cited by examiner]
US 20150344844A1 · Better et al. · 2015 [cited by applicant]
US 20160122782A1 · Crisman et al. · 2016 [cited by applicant]
US 20160319234A1 · Song · 2016 [cited by examiner]
US 20170037369A1 · Ramsborg et al. · 2017 [cited by applicant]
US 20170051252A1 · Morgan et al. · 2017 [cited by applicant]
US 20190211294A1 · Karnieli · 2019 [cited by applicant]
US 20190249130A1 · Griffin et al. · 2019 [cited by applicant]
US 20190358633A1 · Collins · 2019 [cited by examiner]
US 20200208095A1 · Oram · 2020 [cited by examiner]
US 20210147785A1 · Buzalewicz · 2021 [cited by examiner]
AU 2002324169A1 · 2003 [cited by applicant]
DE 4021123A1 · 1991 [cited by applicant]
EP 0248675A1 · 1987 [cited by applicant]
GB 1356794A · 1974 [cited by applicant]
JP 2119772A · 1990 [cited by applicant]
JP 2174848A · 1990 [cited by applicant]
JP 3500847A · 1991 [cited by applicant]
JP 5503418A · 1993 [cited by applicant]
JP 654678A · 1994 [cited by applicant]
JP 6261736A · 1994 [cited by applicant]
JP 7501206A · 1995 [cited by applicant]
JP H0856646A · 1996 [cited by applicant]
JP H11507229A · 1999 [cited by applicant]
JP 2001275659A · 2001 [cited by applicant]
JP 2001517428A · 2001 [cited by applicant]
JP 2002500004A · 2002 [cited by applicant]
JP 2004147555A · 2004 [cited by applicant]
KR 200243145Y1 · 2001 [cited by applicant]
WO 9105849A1 · 1991 [cited by applicant]
WO 9303142A1 · 1993 [cited by applicant]
WO 199712960A2 · 1997 [cited by applicant]
WO 9933951A1 · 1999 [cited by applicant]
WO 9947922A2 · 1999 [cited by applicant]
WO 2000046349A1 · 2000 [cited by applicant]
WO 0102030A2 · 2001 [cited by applicant]
WO 2001000783A2 · 2001 [cited by applicant]
WO 2002028996A1 · 2002 [cited by applicant]
WO 02088295A1 · 2002 [cited by applicant]
WO 03022985A2 · 2003 [cited by applicant]
WO 03087292A2 · 2003 [cited by applicant]
WO 2003085101A1 · 2003 [cited by applicant]
WO 2013151755A1 · 2013 [cited by applicant]
WO 2015162211A1 · 2015 [cited by applicant]
WO 2016069993A1 · 2016 [cited by applicant]
WO 2016118780A1 · 2016 [cited by applicant]
WO 2016168275A1 · 2016 [cited by applicant]
WO 2017068425A1 · 2017 [cited by applicant]
WO 2018015561A1 · 2018 [cited by applicant]
WO 2018136566A1 · 2018 [cited by applicant]
WO 2019046766A2 · 2019 [cited by applicant]
Konstantin B. Konstantinov “Monitoring and Control of the Physiological State of Cell Cultures” Biotechnology and Bioengineering, vol. 52, pp. 271-289 (1996) (Year: 1996). [cited by applicant]
Farndale “Pulsed Electromagnetic Fields Promote Collagen Production in Bone Marrow Fibroblasts via Athermal Mechanisms” Calcif Tissue Int (1985) 37:178-182. [cited by applicant]
Shi et al., Performance of Mammalian Cell Culture Bioreactor with a New Impeller Design, Biotechnology and Bioengineering, Jun. 20, 1992, pp. 260-270, vol. 40, John Wiley & Sons, Inc. [cited by applicant]
Declaration from Mark Selker, Submitted in [cited by applicant]
Declaration from James C. Leung, Submitted in [cited by applicant]
Aitken-Christie et al., Automation in Plant tissue culture—general introduction and overview, in Automation and Environmental Control in Plant Tissue Culture 757 (J. Aitken-Christie, T. Kozai & M. Lila Smith eds., 1995). [cited by applicant]
Apel et al., Integrated Clinical Scale Manufacturing System for Cellular Products Derived by Magnetic Cell Separation, Centrifugation and Cell Culture, Chemie Ingenieur Technik (2013). [cited by applicant]
Armstrong et al., Clinical Systems for the Production of Cells and Tissues for Human Therapy, in Novel Therapeutics From Modern Biotechnology 221 (D.L. Oxender et al. eds., 1999). [cited by applicant]
Blaeschke et al., Induction of A Central Memory and Stem Cell Memory Phenotype in Functionally Active CD4+ and CD8+ CAR T Cells Produced in an Automated Good Manufacturing Practice System for the Treatment of CD19+ Acut… [cited by applicant]
Bohnenkamp et al., Bioprocess development for the cultivation of human T-lymphocytes in a clinical scale, Cytotechnology (2002). [cited by applicant]
Bousso, T-cell activation by dendritic cells in the lymph node: lessons from the movies, 8 Nature Reviews Immunology 675 (2008) (“Bousso 2008”). [cited by applicant]
Basic and Clinical Immunology: Antigen Presentation, T Cell Activation and Deactivation, CLEVELANDCLINICCME, found at https://www.youtube.com/watch?v=EfYpkA4AmFo (2017), last visited Dec. 6, 2020 (“Cleveland Clinic vide… [cited by applicant]
Kaiser et al., Towards a Commercial Process for the Manufacture of Genetically Modified T Cells for Therapy, 22 Cancer Gene Therapy 72-78 (2015). [cited by applicant]
Kempner et al., A Review of Cell Culture Automation, 7 Journal of the Association for Laboratory Automation 56 (2002) (“Kempner 2002”). [cited by applicant]
Koller et al., Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system, Bone Marrow Transplantation (1998) (“Koller 1998”). [cited by applicant]
Koller et al., Large-Scale Expansion of Human Stem and Progenitor Cells from Bone Marrow Mononuclear Cells in Continuous Perfusion Cultures, Blood (1993) (“Koller 1993A”). [cited by applicant]
Koller et al., Tissue Engineering: Reconstitution of Human Hematopoiesis Ex Vivo, Biotechnology and Bioengineering (1993) (“Koller 1993B”). [cited by applicant]
Kostov et al., Low-Cost Microbioreactor for High-Throughput Bioprocessing, 72 Biotechnology and Bioengineering, Feb. 5, 2001 (“Kostov 2001”). [cited by applicant]
Krug et al., A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor, Cancer Immunol Immunotherapy (2014) (“Krug 2014”). [cited by applicant]
Levine et al., Global Manufacturing of CAR T Cell Therapy, 4 Molecular Therapy: Methods & Clinical Development 92 (2017). [cited by applicant]
Lock et al., Automated Manufacturing of Potent CD20-Directed Chimeric Antigen Receptor T Cells for Clinical Use, 28 Human Gene Therapy 10 (2017), (“Lock 2017”). [cited by applicant]
Lu et al., A Rapid Cell Expansion Process for Production of Engineered Autologous CART Cell Therapies, 27 Human Gene Therapy 6 (2016). [cited by applicant]
Mock et al., Automated manufacturing of chimeric antigen receptor T cells for adoptive immunotherapy using CliniMACS Prodigy, Cytotherapy (2016). [cited by applicant]
Morse, Technology evaluation: Stem-cell therapy, Aastrom Biosciences Inc., Current Opinion in Molecule Therapeutics (1999) (“Morse 1999”). [cited by applicant]
Oh et al., Frequent Harvesting from Perfused Bone Marrow Cultures Results in Increased Overall Cell and Progenitor Expansion, Biotechnology and Bioengineering (1994). [cited by applicant]
Priesner et al., Automated Enrichment, Transduction, and Expansion of Clinical-Scale CD62L+ T Cells for Manufacturing of Gene Therapy Medicinal Products, 27 Human Gene Therapy 10, 860-869 (2016). [cited by applicant]
Rosazza et al., Gene Electrotransfer: A Mechanistic Perspective, Current Gene Therapy (2016) (“Rosazza 2016”). [cited by applicant]
Shi et al., “Performance of Mammalian Cell Culture Bioreactor with a New Impeller Design” Biotechnology and Bioengineering, vol. 40, pp. 260-270 (1992). [cited by applicant]
Stiff et al., Autologous transplantation of ex vivo expanded bone marrow cells grown from small aliquots after high-dose chemotherapy for breast cancer, Blood (2000) (“Stiff 2000”). [cited by applicant]
Wang et al., Clinical Manufacturing of CAR T Cells: Foundation of a Promising Therapy, 3 Molecular Therapy—Oncolytics 1 (2016). [cited by applicant]
Wang et al., Manufacture of Tumor- and Virus-specific T Lymphocytes for Adoptive Cell Therapies, 22 Cancer Gene Therapy 2 (2015). [cited by applicant]
Zhang et al., Characterization of clinical grade CD19 chimeric antigen receptor T cells produced using automated CliniMACS Prodigy system, Drug Design, Development and Therapy (2018) (“Zhang 2018”). [cited by applicant]
Zhu et al., Closed-system manufacturing of CD19 and dual-targeted CD20/19 chimeric antigen receptor T cells using the CliniMACS Prodigy device at an academic medical center, Cytotherapy (2018). [cited by applicant]