IP Library Granted Patent US 12,545,873
Granted Patent B2
US 12,545,873 · App. 17/732,239 · Granted Feb 10, 2026

Self-enclosed bioreactor for vascularized tissue constructs

Inventors: Kevin Daniel Roehm (Huntsville, AL); Carrie Lynn German (Huntsville, AL); Balabhaskar Prabhakarpandian (Madison, AL); Wesley David Grove (Huntsville, AL)
Assignee: CFD RESEARCH CORPORATION
C12M21/08C12M23/30C12N5/0018C12N5/0062C12N5/0068C12M23/40C12M27/18C12N2533/90
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Quick Facts
Patent No.
US 12,545,873
App. No.
17/732,239
Granted
Feb 10, 2026
Kind
B2
Abstract

A bioreactor can include: a tissue culture chamber; at least one inlet port into the tissue culture chamber; an inlet port member located in each inlet including an inlet tube extending into the tissue culture chamber; at least one outlet port into the tissue culture chamber; an outlet port member located in each outlet including an outlet tube extending into the tissue culture chamber; an optical cover; and a hydrogel can be located in the tissue culture chamber having at least one lumen fluidly coupling the inlet tube to the outlet tube, wherein an inlet interface region of the hydrogel is constrained around the inlet tube and an outlet interface region of the hydrogel is constrained around the outlet tube.

Claims (62)

1 . A tissue culture device comprising:

a body defining a tissue culture chamber having a primary chamber cavity, an inlet cavity, and an outlet cavity;

at least one inlet port through the body into the inlet cavity of the tissue culture chamber;

an inlet port member located in each inlet port, each inlet port member including an inlet tube extending into the inlet cavity;

at least one outlet port through the body into the outlet cavity of the tissue culture chamber;

an outlet port member located in each outlet port, each outlet port member including an outlet tube extending into the outlet cavity; and

an optical cover coupled with the body to define a wall of the primary chamber cavity of the tissue culture chamber, wherein the optical cover is optically transmissive.

2 . The tissue culture device of claim 1 , wherein the body includes an inlet shoulder containing the inlet cavity, an outlet shoulder containing the outlet cavity, each inlet port is through the inlet shoulder, and each outlet port is through the outlet shoulder.

3 . The tissue culture device of claim 1 , wherein the optical cover is shaped as a lid that is removably couplable with the body.

4 . The tissue culture device of claim 3 , wherein the lid is threadedly coupled with the body.

5 . The tissue culture device of claim 3 , wherein the lid includes a viewing recess having a bottom wall being optically transmissive into the tissue culture chamber.

6 . The tissue culture device of claim 1 , further comprising a hydrogel in the tissue culture chamber, inlet cavity, and outlet cavity, so as to be around the inlet tube and outlet tube.

7 . The tissue culture device of claim 6 , wherein the inlet cavity is configured to constrain the hydrogel around the inlet tube and the outlet cavity is configured to constrain the hydrogel around the outlet tube.

8 . The tissue culture device of claim 7 , further comprising a fluid tight seal at an interface of the hydrogel and the inlet tube and a fluid tight seal at an interface of the hydrogel and the outlet tube.

9 . The tissue culture device of claim 6 , further comprising at least one lumen in the hydrogel fluidly coupling the inlet tube and the outlet tube.

10 . A bioreactor comprising:

a body defining a tissue culture chamber;

at least one inlet port through the body into the tissue culture chamber;

an inlet port member located in each inlet port, each inlet port member including an inlet tube extending into the tissue culture chamber;

at least one outlet port through the body into the tissue culture chamber;

an outlet port member located in each outlet port, each outlet port member including an outlet tube extending into the tissue culture chamber;

an optical cover coupled with the body to define a wall of the primary chamber cavity of the tissue culture chamber, wherein the optical cover is optically transmissive; and

a hydrogel in the tissue culture chamber having at least one lumen fluidly coupling the inlet tube to the outlet tube, wherein an inlet interface region of the hydrogel is constrained around the inlet tube and an outlet interface region of the hydrogel is constrained around the outlet tube.

11 . The bioreactor of claim 10 , wherein the tissue culture chamber has a primary chamber cavity, an inlet cavity, and an outlet cavity, and:

each inlet port is through the body into the inlet cavity of the tissue culture chamber;

each inlet port member includes the inlet tube extending into the inlet cavity;

each outlet port is through the body into the outlet cavity of the tissue culture chamber; and

each outlet port member includes the outlet tube extending into the outlet cavity.

12 . The bioreactor of claim 11 , wherein the body includes an inlet shoulder containing the inlet cavity, an outlet shoulder containing the outlet cavity, each inlet port is through the inlet shoulder, and each outlet port is through the outlet shoulder.

13 . The bioreactor of claim 12 , wherein the optical cover is shaped as a lid that is removably couplable with the body, and wherein the lid includes a viewing recess having a bottom wall being optically transmissive into the tissue culture chamber.

14 . The bioreactor of claim 10 , further comprising a fluid tight seal at an interface of the hydrogel and the inlet tube and a fluid tight seal at an interface of the hydrogel and the outlet tube.

15 . The bioreactor of claim 10 , further comprising cells in the hydrogel.

16 . The bioreactor of claim 10 , further comprising vascular endothelial cells on lumen walls of the lumen.

17 . The bioreactor of claim 16 , further comprising tissue cells in the hydrogel.

18 . The bioreactor of claim 15 , wherein the cells are stem cells, red blood cells, white blood cells, platelet cells, epithelial cells, nerve cells, muscle cells, cartilage cells, bone cells, connective tissue cells, skin cells, endothelial cells, fat cells, sex cells, organ cells, or combinations thereof.

19 . The bioreactor of claim 18 , wherein the at least one lumen is configured in diameter and spacing based on an organ, such that oxygen and/or nutrients diffuse from media in the at least one lumen to the cells in the organ in the hydrogel.

20 . The bioreactor of claim 10 , wherein the at least one lumen includes a sacrificial material therein.

21 . A bioreactor system comprising:

the bioreactor of claim 10 ; and

an imaging system having at least one camera optically coupled with the tissue culture chamber.

22 . A bioreactor system comprising:

the bioreactor of claim 10 ; and

a pressure system fluidly coupled with the inlet tube and/or outlet tube.

23 . The bioreactor system of claim 22 , further comprising a media system comprising a media for flowing through the at least one lumen.

24 . The bioreactor system of claim 23 , further comprising a collection system configured for receiving output media from the outlet tube.

25 . A bioreactor system comprising:

the tissue culture device of claim 10 ; and

an analytical system configured to analyze output samples from the outlet tube.

26 . A method of manufacturing a bioreactor comprising:

providing the tissue culture device of claim 1 ;

forming a hydrogel in the tissue culture chamber that has at least one lumen fluidly coupling the inlet tube to the outlet tube of the tissue culture device, wherein an inlet interface region of the hydrogel is constrained around the inlet tube and an outlet interface region of the hydrogel is constrained around the outlet tube.

27 . The method of claim 26 , further comprising:

forming a sacrificial material in the hydrogel in the shape of the at least one lumen; and

removing the sacrificial material from the hydrogel to result in the at least one lumen being fluidly coupled with the inlet tube and outlet tube.

28 . The method of claim 27 , further comprising:

printing the sacrificial material on a portion of the hydrogel; and

forming the hydrogel with the sacrificial material therein.

29 . The method of claim 26 , further comprising:

determining a type of tissue for a tissue construct;

obtaining a design for the at least one lumen in the hydrogel for the tissue construct based on the type of tissue; and

forming the hydrogel to include the design for the at least one lumen in the hydrogel.

30 . The method of claim 29 , wherein the design includes a diameter of each lumen and spacing between a plurality of the lumen.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 28, 2022
From: ROEHM, KEVIN DANIEL; GERMAN, CARRIE LYNN; PRABHAKARPANDIAN, BALABHASKAR; GROVE, WESLEY DAVID
To: CFD RESEARCH CORPORATION
Reel/Frame 059759/0971 →
Continuity (1)
Related Publication 20230348830A1 · Nov 2, 2023
References Cited (49)
US 9023642B2 · Kleis et al. · 2015 [cited by applicant]
US 9758762B2 · Meiron et al. · 2017 [cited by applicant]
US 9783768B2 · Larcher et al. · 2017 [cited by applicant]
US 9902929B2 · Wu · 2018 [cited by applicant]
US 10214714B2 · Maria de Peppo · 2019 [cited by applicant]
US 10214718B2 · Berteau et al. · 2019 [cited by applicant]
US 20080261306A1 · Neumann · 2008 [cited by examiner]
US 20080311650A1 · Jakob · 2008 [cited by examiner]
US 20130143230A1 · Tolias · 2013 [cited by examiner]
US 20130344531A1 · Akra · 2013 [cited by examiner]
US 20150212071A1 · Berry et al. · 2015 [cited by applicant]
US 20160130543A1 · Daniele · 2016 [cited by examiner]
US 20170096627A1 · Smith et al. · 2017 [cited by applicant]
US 20180030409A1 · Lewis et al. · 2018 [cited by applicant]
US 20180066220A1 · Nath · 2018 [cited by examiner]
US 20190022283A1 · Lewis et al. · 2019 [cited by applicant]
US 20190225925A1 · Vunjak-Novakovic et al. · 2019 [cited by applicant]
US 20200088719A1 · Gevaert et al. · 2020 [cited by applicant]
US 20200181556A1 · Eberth et al. · 2020 [cited by applicant]
US 20200332242A1 · Russell · 2020 [cited by applicant]
US 20240026260A1 · Takeuchi · 2024 [cited by examiner]
H. Wobma, G. Vunjak-Novakovic; “Tissue Engineering and Regenerative Medicine 2015: A Year in Review”; Tissue Engineering—Part B: Reviews; Apr. 2016; vol. 22(2); pp. 101-113; https://doi.org/10.1089/ten.teb.2015.0535; Ep… [cited by applicant]
I. Sukmana; “Microvascular guidance: A challenge to support the development of vascularised tissue engineering construct”; The Scientific World Journal; Apr. 2012; vol. 2012(5110); https://doi.org/10.1100/2012/201352. [cited by applicant]
M. Lovett, K. Lee, A. Edwards, D.L. Kaplan; “Vascularization strategies for tissue engineering”; Tissue Engineering - Part B: Reviews; vol. 15(3); Jul. 2009; pp. 353-370; https://doi.org/10.1089/ten.teb.2009.0085. [cited by applicant]
D.B. Kolesky, K.A. Homan, M.A. Skylar-Scott, J.A. Lewis; “Three-dimensional bioprinting of thick vascularized tissues”; Proceedings of the National Academy of Sciences of the U.S.A; Mar. 22, 2016; 113(12); pp. 3179-3184… [cited by applicant]
B. Grigoryan, S.J. Paulsen, D.C. Corbett, D.W. Sazer, C.L. Fortin, A.J. Zaita, P.T. Greenfield, N.J. Calafat, J.P. Gounley, A.H. Ta, F. Johansson, A. Randles, J.E. Rosenkrantz, J.D. Louis-Rosenberg, P.A. Galie, K.R. Ste… [cited by applicant]
D.B. Kolesky, R.L. Truby, A.S. Gladman, T.A. Busbee, K.A. Homan, J.A. Lewis; “3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs”; Advanced Materials; May 21, 2014; vol. 26(19); pp. 3124-3130; ht… [cited by applicant]
J.S. Miller, K.R. Stevens, M.T. Yang, B.M. Baker, D.H.T. Nguyen, D.M. Cohen, E. Toro, A.A. Chen, P.A. Galie, X. Yu, R. Chaturvedi, S.N. Bhatia, C.S. Chen; “Rapid casting of patterned vascular networks for perfusable eng… [cited by applicant]
M.A. Skylar-Scott, S.G.M. Uzel, L.L. Nam, J.H. Ahrens, R.L. Truby, S. Damaraju, J.A. Lewis; “Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels”; Science Advances. Sep. … [cited by applicant]
R.H. Wenger, V. Kurtcuoglu, C.C. Scholz, H.H. Marti, D. Hoogewijs; “Frequently asked questions in hypoxia research”; Hypoxia (Auckl). Sep. 18, 2015; vol. 3(35-43); https://doi.org/10.2147/HP.S92198; PMID: 27774480; PMCI… [cited by applicant]
C.A. V Rodrigues, T.G. Fernandes, M.M. Diogo, C. Lobato Da Silva, J.M.S. Cabral; “Stem cell cultivation in bioreactors”; Biotechnology Advances; Nov.-Dec. 2011; vol. 29(6); pp. 815-829; https://doi.org/10.1016/j.biotech… [cited by applicant]
J.A. King, W.M. Miller; “Bioreactor Development for Stem Cell Expansion and Controlled Differentiation”; Current Opinion in Chemical Biology; Aug. 11, 2007; vol. 4; pp. 394-398; https://doi.org/10.1016/J.CBPA.2007.05.03… [cited by applicant]
K.M. Panchalingam, S. Jung, L. Rosenberg, L.A. Behie; “Bioprocessing strategies for the large-scale production of human mesenchymal stem cells: a review”; Stem Cell Research & Therapy; Nov. 23, 2015; vol. 6:(225) pp. 1-… [cited by applicant]
M.J. Powers, K. Domansky, M.R. Kaazempur-Mofrad, A. Kalezi, A. Capitano, A. Upadhyaya, p. Kurzawski, K.E. Wack, D.B. Stolz, R. Kamm, L.G. Grifith; “A Microfabricated Array Bioreactor for Perfused 3D Liver Culture”; Biot… [cited by applicant]
Egger D, Spitz S, Fischer M, Handschuh S, Glösmann M, Friemert B, Egerbacher M, Kasper C.; “Application of a Parallelizable Perfusion Bioreactor for Physiologic 3D Cell Culture”; Cells Tissues Organs; 2017; vol. 203(5):… [cited by applicant]
S.M. Warren, A.M. Sailon, A.C. Allori, E.H. Davidson, D.D. Reformat, R.J. Allen; “A novel flow-perfusion bioreactor supports 3D dynamic cell culture”; Journal of Biomedicine and Biotechnology; vol. 2009; Dec. 9, 2009; h… [cited by applicant]
C.L. German, S. V. Madihally; “Applications of Computational Modelling and Simulation of Porous Medium in Tissue Engineering”; Computation; vol. 4(1); p. 7; Feb. 6, 2016; https://doi.org/10.3390/COMPUTATION4010007. [cited by applicant]
C.C. Michel; “Starling: the formulation of his hypothesis of microvascular fluid exchange and its significance after 100 years”; Experimental Physiology; Translation and Integration. vol. 82(1) Jan. 1997; pp. 1-30; doi:… [cited by applicant]
R. McMurtrey; Analytic models of oxygen and nutrient diffusion, metabolism dynamics, and architecture optimization in three-dimensional tissue constructs with applications and Insights in Cerebral Organoids; .-Tissue En… [cited by applicant]
C. Wang, H. Lu, M.A, Schwartz; “A novel in vitro flow system for changing flow direction on endothelial cells”; Journal of Biomechanics,; vol. 45(7); 2012; pp. 1212-1218; https://doi.org/10.1016/j.jbiomech.2012.01.045. [cited by applicant]
P.M. Hinderliter, K.R. Minard, G. Orr, W.B. Chrisler, B.D. Thrall, J.G. Pounds, J.G. Teeguarden; “ISDD: A computational model of particle sedimentation, diffusion and target cell dosimetry for in vitro toxicity studies”… [cited by applicant]
S. Pradhan, I. Hassani, I, Seeto WJ, Lipke EA; “PEG-fibrinogen hydrogels for three-dimensional breast cancer cell culture”; Whitepaper; Wiley Online Library; published Oct. 14, 2016; 17 pages; DOI:10.1002/jbm.a.35899. [cited by applicant]
M. Radisic, J. Malda, E. Epping, W. Geng, R. Langer, G. Vunjak-Novakovic; “Oxygen Gradients Correlate with Cell Density and Cell Viability in Engineered Cardiac Tissue”; Whitepaper; Willey InterScience; published Nov. 3… [cited by applicant]
K. Sekine, Y. Kagawa, E. Maeyama, H. Ota, Y. Haraguchi, K. Matsuura, T. Shimizu; “Oxygen consumption of human heart cells in monolayer culture”; Biochemical and Biophysical Research Communications; 452(3); Sep. 26, 2014… [cited by applicant]
C. Magliaro, G. Mattei, F. Iacoangeli, A. Corti, V. Piemonte, A. Ahluwalia; “Oxygen Consumption Characteristics in 3D Constructs Depend on Cell Density”; Frontiers in Bioengineering and Biotechnology; vol. 7, Issue 251;… [cited by applicant]
M.K. Gelber, G. Hurst, T.J. Comi, R. Bhargava; Model-guided design and characterization of a high-precision 3D printing process for carbohydrate glass:; Additive Manufacturing; vol. 22; Aug. 2018; pp. 38-50. https://doi… [cited by applicant]
M.K. Gelber, R. Bhargava; “Monolithic multilayer microfluidics via sacrificial molding of 3D-printed isomalt”; Lab on a Chip; vol. 15, Issue 7; Apr. 7, 2015; pp. 1736-1741. [cited by applicant]
Kevin D. Roehm, Sundararajan V. Madihally; “Bioprinted chitosan-gelatin thermosensitive hydrogels using an inexpensive 3D printer”; Biofabrication; Nov. 30, 2017; 10(1):015002. [cited by applicant]
A. Przekwas, T. Friend, R. Teixeira, Z. Chen, P. Wilkerson; “Spatial modeling tools for cell biology”, Final Technical Report; Oct. 2006; located at: https://apps.dtic.mil/sti/citations/ADA460852; 115 pages. [cited by applicant]