Bioprocessing of human mesenchymal stem/stromal cells for therapeutic use: Current technologies and challenges

Biochemical Engineering Journal

Volumn 108, Issue , 2016, Pages 3-13

  Schnitzler, Aletta C ^a   Verma, Anjali ^a   Kehoe, Daniel E ^a   Jing, Donghui ^a   Murrell, Julie R ^a   Der, Kara A ^a   Aysola, Manjula ^a   Rapiejko, Peter J ^a   Punreddy, Sandhya ^a   Rook, Martha S ^a  

^a EMD Millipore   (United States)

Author keywords

Bioreactors; Cell culture media; Mesenchymal stem stromal cells; Microcarriers; Scale up; Separation

Indexed keywords

ANIMALS; BIOREACTORS; CELL CULTURE; CELLS; CYTOLOGY; SEPARATION; STEM CELLS;

CELL CULTURE MEDIA; CELL CULTURE SUBSTRATE; CELLULAR THERAPEUTICS; DOWNSTREAM-PROCESSING; FUNCTIONAL CHARACTERISTICS; MANUFACTURING PROCESS; MICROCARRIERS; SCALE-UP;

ANIMAL CELL CULTURE;

DISSOLVED OXYGEN; GROWTH FACTOR;

ARTICLE; BIOPROCESS; BIOREACTOR; CELL ADHESION; CELL CULTURE; CELL EXPANSION; CELL LYSATE; CELL SEPARATION; CELLS; CRYOPRESERVATION; DOWNSTREAM PROCESSING; MESENCHYMAL STEM CELL; MESENCHYMAL STROMA CELL; NUTRIENT CONTENT; PRIORITY JOURNAL; STEM CELL TRANSPLANTATION; THROMBOCYTE;

EID: 84959342328     PISSN: 1369703X     EISSN: 1873295X     Source Type: Journal    
DOI: 10.1016/j.bej.2015.08.014     Document Type: Article

References (102)

1. Augello A., Kurth T.B., de Bari C. Mesenchymal stem cells: a perspective from in vitro cultures to in vivo migration and niches. Eur. Cell Mater. 2010, 20:121-133.
2. Chamberlain G., Fox J., Ashton B., Middleton J. Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells 2007, 25:2739-2749.
3. Rowley J., Abraham E., Campbell A., Brandwein H., Oh S. Meeting lot-size challenges of manufacturing adherent cells for therapy. BioProcess Int. 2012, 10:16-22.
4. Simaria A.S., Hassan S., Varadaraju H., Rowley J., Warren K., Vanek P., Farid S.S. Allogeneic cell therapy bioprocess economics and optimization: single-use cell expansion technologies. Biotechnol. Bioeng. 2014, 111:69-83.
5. [Internet]. Bethesda (MD): National Library of Medicine (US). Available from: , 2000 (cited 2015 March 27). http://https://clinicaltrials.gov/ct2/home.
6. Cardio3 BioSciences, Efficacy and safety of bone marrow-derived mesenchymal cardiopoietic cells (C3BS-CQR-1) for the treatment of chronic advanced ischemic heart failure, In: [Internet]. Bethesda (MD): National Library of Medicine (US). Available from: , 2000 (cited 2015 March 17). http://https://clinicaltrials.gov/show/NCT01768702.
7. Osiris announces agreement for culture-expanded MSCs including Prochymal worth up yo $100 million plus royalties, In: [Internet]. Available from: , 2000 (cited 2015 March 27). http://investor.osiris.com/releasedetail.cfm%3FReleaseID=796371.
8. Mesoblast Ltd., A prospective study of remestemcel-L, ex-vivo cultured adult human mesenchymal stromal cells, for the treatment of pediatric patients who have failed to respond to steroid treatment for acute GVHD, In: [Internet]. Bethesda (MD): National Library of Medicine (US). Available from: , 2000 (cited 2015 March 27). http://https://clinicaltrials.gov/show/NCT02336230.
9. Pfizer; Athersys Inc., A phase 2 randomized, double-blind, placebo-controlled, parallel group, multi-center study to investigate the safety and efficacy of Multistem (Pf-05285401) in subjects with moderate to severe ulcerative colitis, In: [Internet]. Bethesda (MD): National Library of Medicine (US). Available from: , 2000 (cited 2015 March 17). http://https://clinicaltrials.gov/show/NCT01240915.
10. Athersys announces results from phase 2 study of MultiStem® cell therapy for ulcerative colitis, In: [Internet]. Available from: , 2000 (cited 2015 March 27). http://www.athersys.com/releasedetail.cfm%3FReleaseID=842936.
11. Athersys Inc., Double-blind, randomized, placebo-controlled phase 2 safety and efficacy trial of MultiStem® in adults with ischemic stroke, In: [Internet]. Bethesda (MD): National Library of Medicine (US). Available from: , 2000 (cited 2015 March 22). http://https://clinicaltrials.gov/show/NCT01436487.
12. Baksh D., Davies J.E., Zandstra P.W. Adult human bone marrow-derived mesenchymal progenitor cells are capable of adhesion-independent survival and expansion. Exp. Hematol. 2003, 31:723-732.
13. Baraniak P.R., McDevitt T.C. Scaffold-free culture of mesenchymal stem cell spheroids in suspension preserves multilineage potential. Cell Tissue Res. 2012, 347:701-711.
14. Alimperti S., Lei P., Wen Y., Tian J., Campbell A.M., Andreadis S.T. Serum-free spheroid suspension culture maintains mesenchymal stem cell proliferation and differentiation potential. Biotechnol. Prog. 2014, 30:974-983.
15. Sensebe L., Bourin P., Tarte K. Good manufacturing practices production of mesenchymal stem/stromal cells. Hum. Gene Ther. 2011, 22:19-26.
16. Fossett E., Khan W.S. Optimising human mesenchymal stem cell numbers for clinical application: a literature review. Stem Cells Int. 2012, 2012:465259.
17. Vidal M.A., Walker N.J., Napoli E., Borjesson D.L. Evaluation of senescence in mesenchymal stem cells isolated from equine bone marrow, adipose tissue, and umbilical cord tissue. Stem Cells Dev. 2012, 21:273-283.
18. Dominici M., Le Blanc K., Mueller I., Slaper-Cortenbach I., Marini F., Krause D., Deans R., Keating A., Prockop D., Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006, 8:315-317.
19. Bourin P., Bunnell B.A., Casteilla L., Dominici M., Katz A.J., March K.L., Redl H., Rubin J.P., Yoshimura K., Gimble J.M. Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy 2013, 15:641-648.
20. Samsonraj R.M., Rai B., Sathiyanathan P., Puan K.J., Rotzschke O., Hui J.H., Raghunath M., Stanton L.W., Nurcombe V., Cool S.M. Establishing criteria for human mesenchymal stem cell potency. Stem Cells 2015, 33:1878-1891.
21. De Ugarte D.A., Alfonso Z., Zuk P.A., Elbarbary A., Zhu M., Ashjian P., Benhaim P., Hedrick M.H., Fraser J.K. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow. Immunol. Lett. 2003, 89:267-270.
22. Tondreau T., Meuleman N., Delforge A., Dejeneffe M., Leroy R., Massy M., Mortier C., Bron D., Lagneaux L. Mesenchymal stem cells derived from CD133-positive cells in mobilized peripheral blood and cord blood: proliferation, Oct4 expression, and plasticity. Stem Cells 2005, 23:1105-1112.
23. Galderisi U., Giordano A. The gap between the physiological and therapeutic roles of mesenchymal stem cells. Med. Res. Rev. 2014, 34:1100-1126.
24. Sun L.Y., Lin S.Z., Li Y.S., Harn H.J., Chiou T.W. Functional cells cultured on microcarriers for use in regenerative medicine research. Cell Transplant. 2011, 20:49-62.
25. Qian L., Saltzman W.M. Improving the expansion and neuronal differentiation of mesenchymal stem cells through culture surface modification. Biomaterials 2004, 25:1331-1337.
26. Morla A., Zhang Z., Ruoslahti E. Superfibronectin is a functionally distinct form of fibronectin. Nature 1994, 367:193-196.
27. Mukhopadhyay A., Mukhopadhyay S.N., Talwar G.P. Influence of serum proteins on the kinetics of attachment of Vero cells to cytodex microcarriers. J. Chem. Technol. Biotechnol. 1993, 56:369-374.
28. Sart S., Errachid A., Schneider Y.J., Agathos S.N. Modulation of mesenchymal stem cell actin organization on conventional microcarriers for proliferation and differentiation in stirred bioreactors. J. Tissue Eng. Regen. Med. 2013, 7:537-551.
29. Santos F., Andrade P.Z., Abecasis M.M., Gimble J.M., Chase L.G., Campbell A.M., Boucher S., Vemuri M.C., Silva C.L., Cabral J.M. Toward a clinical-grade expansion of mesenchymal stem cells from human sources: a microcarrier-based culture system under xeno-free conditions. Tissue Eng. Part C Methods 2011, 17:1201-1210.
30. Mauney J.R., Kaplan D.L., Volloch V. Matrix-mediated retention of osteogenic differentiation potential by human adult bone marrow stromal cells during ex vivo expansion. Biomaterials 2004, 25:3233-3243.
31. Dolley-Sonneville P.J., Romeo L.E., Melkoumian Z.K. Synthetic surface for expansion of human mesenchymal stem cells in xeno-free, chemically defined culture conditions. PLoS One 2013, 8:e70263.
32. Zhao W., Li X., Liu X., Zhang N., Wen X. Effects of substrate stiffness on adipogenic and osteogenic differentiation of human mesenchymal stem cells. Mater. Sci. Eng. C 2014, 40:316-323.
33. McMurray R.J., Gadegaard N., Tsimbouri P.M., Burgess K.V., McNamara L.E., Tare R., Murawski K., Kingham E., Oreffo R.O., Dalby M.J. Nanoscale surfaces for the long-term maintenance of mesenchymal stem cell phenotype and multipotency. Nat. Mater. 2011, 10:637-644.
34. Engler A.J., Sen S., Sweeney H.L., Discher D.E. Matrix elasticity directs stem cell lineage specification. Cell 2006, 126:677-689.
35. Hervy M., Weber J.L., Pecheul M., Dolley-Sonneville P., Henry D., Zhou Y., Melkoumian Z. Long term expansion of bone marrow-derived hMSCs on novel synthetic microcarriers in xeno-free, defined conditions. PLoS One 2014, 9:e92120.
36. Jing D., Sunil N., Punreddy S., Aysola M., Kehoe D., Murrel J., Rook M., Niss K. Growth kinetics of human mesenchymal stem cells in a 3-L single-use, stirred-tank bioreactor. BioPharm Int. 2013, 26:28-38.
37. dos Santos F., Campbell A., Fernandes-Platzgummer A., Andrade P.Z., Gimble J.M., Wen Y., Boucher S., Vemuri M.C., da Silva C.L., Cabral J.M. A xenogeneic-free bioreactor system for the clinical-scale expansion of human mesenchymal stem/stromal cells. Biotechnol. Bioeng. 2014, 111:1116-1127.
38. Goh T.K., Zhang Z.Y., Chen A.K., Reuveny S., Choolani M., Chan J.K., Oh S.K. Microcarrier culture for efficient expansion and osteogenic differentiation of human fetal mesenchymal stem cells. BioRes. Open Access 2013, 2:84-97.
39. Rafiq Q.A., Brosnan K.M., Coopman K., Nienow A.W., Hewitt C.J. Culture of human mesenchymal stem cells on microcarriers in a 5 L stirred-tank bioreactor. Biotechnol. Lett. 2013, 35:1233-1245.
40. Timmins N.E., Kiel M., Gunther M., Heazlewood C., Doran M.R., Brooke G., Atkinson K. Closed system isolation and scalable expansion of human placental mesenchymal stem cells. Biotechnol. Bioeng. 2012, 109:1817-1826.
41. Eibl R., Eibl D. Application of disposable bag bioreactors in tissue engineering and for the production of therapeutic agents. Adv. Biochem. Eng. Biotechnol. 2009, 112:183-207.
42. Chisti Y. Hydrodynamic damage to animal cells. Crit. Rev. Biotechnol. 2001, 21:67-110.
43. Cherry R.S., Papoutsakis E.T. Physical mechanisms of cell damage in microcarrier cell culture bioreactors. Biotechnol. Bioeng. 1988, 32:1001-1014.
44. Chen A.K., Reuveny S., Oh S.K. Application of human mesenchymal and pluripotent stem cell microcarrier cultures in cellular therapy: achievements and future direction. Biotechnol. Adv. 2013, 31:1032-1046.
45. Clark J.M., Hirtenstein M.D. Optimizing culture conditions for the production of animal cells in microcarrier culture. Annals of the New York Academy of Sciences 1981, 369:33-46.
46. Chapman C.M., Nienow A.W., Cooke M., Middleton J.C. Particle-gas-liquid mixing in stirred vessels. Part I: particle-liquid mixing. Chem. Eng. Res. Des. 1983, 61:71-81.
47. Zwietering T.N. Suspending of solid particles in liquid by agitators. Chem. Eng. Sci. 1958, 8:244-253.
48. Ibrahim S., Nienow A.W. Suspension of microcarriers for cell culture with axial flow impellers. Chem. Eng. Res. Des. 2004, 82:1082-1088.
49. Wu S.C., Hsieh W.C., Liau M.Y. Comparisons of microcarrier cell culture processes in one hundred mini-liter spinner flask and fifteen-liter bioreactor cultures. Bioprocess. Eng. 1998, 19:431-434.
50. King J.A., Miller W.M. Bioreactor development for stem cell expansion and controlled differentiation. Curr. Opin. Chem. Biol. 2007, 11:394-398.
51. Croughan M.S., Sayre E.S., Wang D.I. Viscous reduction of turbulent damage in animal cell culture. Biotechnol. Bioeng. 1989, 33:862-872.
52. Hewitt C.J., Lee K., Nienow A.W., Thomas R.J., Smith M., Thomas C.R. Expansion of human mesenchymal stem cells on microcarriers. Biotechnol. Lett. 2011, 33:2325-2335.
53. Godoy-Silva R., Chalmers J.J., Casnocha S.A., Bass L.A., Ma N. Physiological responses of CHO cells to repetitive hydrodynamic stress. Biotechnol. Bioeng. 2009, 103:1103-1117.
54. Kreke M.R., Sharp L.A., Lee Y.W., Goldstein A.S. Effect of intermittent shear stress on mechanotransductive signaling and osteoblastic differentiation of bone marrow stromal cells. Tissue Eng. Part A 2008, 14:529-537.
55. Yeatts A.B., Choquette D.T., Fisher J.P. Bioreactors to influence stem cell fate: augmentation of mesenchymal stem cell signaling pathways via dynamic culture systems. Biochim. Biophys. Acta 2013, 1830:2470-2480.
56. Chow D.C., Wenning L.A., Miller W.M., Papoutsakis E.T. Modeling pO(2) distributions in the bone marrow hematopoietic compartment. I. Krogh's model. Biophys. J. 2001, 81:675-684.
57. Bizzarri A., Koehler H., Cajlakovic M., Pasic A., Schaupp L., Klimant I., Ribitsch V. Continuous oxygen monitoring in subcutaneous adipose tissue using microdialysis. Anal. Chim. Acta 2006, 573-574:48-56.
58. Harrison J.S., Rameshwar P., Chang V., Bandari P. Oxygen saturation in the bone marrow of healthy volunteers. Blood 2002, 99:394.
59. Lavrentieva A., Majore I., Kasper C., Hass R. Effects of hypoxic culture conditions on umbilical cord-derived human mesenchymal stem cells. Cell Commun. Signal. 2010, 8:18.
60. Lonne M., Lavrentieva A., Walter J.G., Kasper C. Analysis of oxygen-dependent cytokine expression in human mesenchymal stem cells derived from umbilical cord. Cell Tissue Res. 2013, 353:117-122.
61. Haque N., Rahman M.T., Abu Kasim N.H., Alabsi A.M. Hypoxic culture conditions as a solution for mesenchymal stem cell based regenerative therapy. Sci. World J. 2013, 2013:632972.
62. Sart S., Agathos S.N., Li Y. Process engineering of stem cell metabolism for large scale expansion and differentiation in bioreactors. Biochem. Eng. J. 2014, 84:74-82.
63. Grayson W.L., Zhao F., Izadpanah R., Bunnell B., Ma T. Effects of hypoxia on human mesenchymal stem cell expansion and plasticity in 3D constructs. J. Cell. Physiol. 2006, 207:331-339.
64. Rauch C., Feifel E., Amann E.M., Spotl H.P., Schennach H., Pfaller W., Gstraunthaler G. Alternatives to the use of fetal bovine serum: human platelet lysates as a serum substitute in cell culture media. Altex 2011, 28:305-316.
65. van der Valk J., Brunner D., De Smet K., Fex Svenningsen A., Honegger P., Knudsen L.E., Lindl T., Noraberg J., Price A., Scarino M.L., Gstraunthaler G. Optimization of chemically defined cell culture media-replacing fetal bovine serum in mammalian in vitro methods. Toxicol. In Vitro 2010, 24:1053-1063.
66. Brindley D.A., Davie N.L., Culme-Seymour E.J., Mason C., Smith D.W., Rowley J.A. Peak serum: implications of serum supply for cell therapy manufacturing. Regen. Med. 2012, 7:7-13.
67. Bernardo M.E., Avanzini M.A., Perotti C., Cometa A.M., Moretta A., Lenta E., Del Fante C., Novara F., de Silvestri A., Amendola G., Zuffardi O., Maccario R., Locatelli F. Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches: further insights in the search for a fetal calf serum substitute. J. Cell. Physiol. 2007, 211:121-130.
68. Iudicone P., Fioravanti D., Bonanno G., Miceli M., Lavorino C., Totta P., Frati L., Nuti M., Pierelli L. Pathogen-free, plasma-poor platelet lysate and expansion of human mesenchymal stem cells. J. Transl. Med. 2014, 12:28.
69. Fekete N., Gadelorge M., Furst D., Maurer C., Dausend J., Fleury-Cappellesso S., Mailander V., Lotfi R., Ignatius A., Sensebe L., Bourin P., Schrezenmeier H., Rojewski M.T. Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components. Cytotherapy 2012, 14:540-554.
70. Doucet C., Ernou I., Zhang Y., Llense J.R., Begot L., Holy X., Lataillade J.J. Platelet lysates promote mesenchymal stem cell expansion: a safety substitute for animal serum in cell-based therapy applications. J. Cell. Physiol. 2005, 205:228-236.
71. Ben Azouna N., Jenhani F., Regaya Z., Berraeis L., Ben Othman T., Ducrocq E., Domenech J. Phenotypical and functional characteristics of mesenchymal stem cells from bone marrow: comparison of culture using different media supplemented with human platelet lysate or fetal bovine serum. Stem Cell Res. Ther. 2012, 3:6.
72. Castegnaro S., Chieregato K., Maddalena M., Albiero E., Visco C., Madeo D., Pegoraro M., Rodeghiero F. Effect of platelet lysate on the functional and molecular characteristics of mesenchymal stem cells isolated from adipose tissue. Curr. Stem Cell Res. Ther. 2011, 6:105-114.
73. Reinhardt J., Stuhler A., Blumel J. Safety of bovine sera for production of mesenchymal stem cells for therapeutic use. Hum. Gene Ther. 2011, 22:775.
74. Jung S., Panchalingam K.M., Rosenberg L., Behie L.A. Ex vivo expansion of human mesenchymal stem cells in defined serum-free media. Stem Cells Int. 2012, 2012:123030.
75. Ludwig T.E., Levenstein M.E., Jones J.M., Berggren W.T., Mitchen E.R., Frane J.L., Crandall L.J., Daigh C.A., Conard K.R., Piekarczyk M.S., Llanas R.A., Thomson J.A. Derivation of human embryonic stem cells in defined conditions. Nat. Biotechnol. 2006, 24:185-187.
76. Jeon M.K., Lim J.B., Lee G.M. Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design. BMC Biotechnol. 2010, 10:70.
77. Lee M.Y., Ryu J.M., Lee S.H., Park J.H., Han H.J. Lipid rafts play an important role for maintenance of embryonic stem cell self-renewal. J. Lipid Res. 2010, 51:2082-2089.
78. Li H., Guo H., Li H. Cholesterol loading affects osteoblastic differentiation in mouse mesenchymal stem cells. Steroids 2013, 78:426-433.
79. Greenberg-Ofrath N., Terespolosky Y., Kahane I., Bar R. Cyclodextrins as carriers of cholesterol and fatty acids in cultivation of mycoplasmas. Appl. Environ. Microbiol. 1993, 59:547-551.
80. dos Santos C., Buera M.P., Mazzobre M.F. Phase solubility studies and stability of cholesterol/beta-cyclodextrin inclusion complexes. J. Sci. Food Agric. 2011, 91:2551-2557.
81. Seamans T.C., Gould S.L., DiStefano D.J., Silberklang M., Robinson D.K. Use of lipid emulsions as nutritional supplements in mammalian cell culture. Ann. N. Y. Acad. Sci. 1994, 745:240-243.
82. Bryan N., Andrews K.D., Loughran M.J., Rhodes N.P., Hunt J.A. Elucidating the contribution of the elemental composition of fetal calf serum to antigenic expression of primary human umbilical-vein endothelial cells in vitro. Biosci. Rep. 2011, 31:199-210.
83. Mafi P., Hindocha S., Mafi R., Griffin M., Khan W.S. Adult mesenchymal stem cells and cell surface characterization-a systematic review of the literature. Open Orthop. J. 2011, 5:253-260.
84. Eto N., Yamada K., Shito T., Shirahata S., Murakami H. Development of a protein-free medium with ferric citrate substituting transferrin for the cultivation of mouse-mouse hybridomas. Agric. Biol. Chem. 1991, 55:863-865.
85. Lu W., Zhao M., Rajbhandary S., Xie F., Chai X., Mu J., Meng J., Liu Y., Jiang Y., Xu X., Meng A. Free iron catalyzes oxidative damage to hematopoietic cells/mesenchymal stem cells in vitro and suppresses hematopoiesis in iron overload patients. Eur. J. Haematol. 2013, 91:249-261.
86. Chase L.G., Lakshmipathy U., Solchaga L.A., Rao M.S., Vemuri M.C. A novel serum-free medium for the expansion of human mesenchymal stem cells. Stem Cell Res. Ther. 2010, 1:8.
87. Jung S., Sen A., Rosenberg L., Behie L.A. Identification of growth and attachment factors for the serum-free isolation and expansion of human mesenchymal stromal cells. Cytotherapy 2010, 12:637-657.
88. Chen A.K., Chew Y.K., Tan H.Y., Reuveny S., Weng Oh S.K. Increasing efficiency of human mesenchymal stromal cell culture by optimization of microcarrier concentration and design of medium feed. Cytotherapy 2015, 17:163-173.
89. McGrail D.J., Ghosh D., Quach N.D., Dawson M.R. Differential mechanical response of mesenchymal stem cells and fibroblasts to tumor-secreted soluble factors. PloS one 2012, 7:e33248.
90. Ponte A.L., Marais E., Gallay N., Langonne A., Delorme B., Herault O., Charbord P., Domenech J. The in vitro migration capacity of human bone marrow mesenchymal stem cells: comparison of chemokine and growth factor chemotactic activities. Stem Cells 2007, 25:1737-1745.
91. D.J. Giard, W.S. Hu, D.I.C. Wang, Detachment of anchorage-dependent cells from microcarriers, European Patent Office (EPO), patent # WO1986001531 A1, Massachusetts Institute of Technology, 1986, 2015.
92. Nienow A.W., Rafiq Q.A., Coopman K., Hewitt C.J. A potentially scalable method for the harvesting of hMSCs from microcarriers. Biochem. Eng. J. 2014, 85:79-88.
93. Ikebe C., Suzuki K. Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols. Biomed. Res Int 2014, 2014:951512.
94. Duffy C.R., Zhang R., How S.E., Lilienkampf A., De Sousa P.A., Bradley M. Long term mesenchymal stem cell culture on a defined synthetic substrate with enzyme free passaging. Biomaterials 2014, 35:5998-6005.
95. Yang H.S., Jeon O., Bhang S.H., Lee S.H., Kim B.S. Suspension culture of mammalian cells using thermosensitive microcarrier that allows cell detachment without proteolytic enzyme treatment. Cell Transplant. 2010, 19:1123-1132.
96. Clarke D., Harati D., Martin J., Rowley J., Keller J., McCaman M., Carrion M., Karnieli O., Maziarz R., Perry R., Oh S., Stanton J. Managing particulates in cellular therapy. Cytotherapy 2012, 14:1032-1040.
97. Frauenschuh S., Reichmann E., Ibold Y., Goetz P.M., Sittinger M., Ringe J. A microcarrier-based cultivation system for expansion of primary mesenchymal stem cells. Biotechnol. Prog. 2007, 23:187-193.
98. Weber C., Pohl S., Portner R., Wallrapp C., Kassem M., Geigle P., Czermak P. Expansion and harvesting of hMSC-TERT. Open Biomed. Eng. J. 2007, 1:38-46.
99. Ullah I., Subbarao R.B., Rho G.J. Human mesenchymal stem cells-current trends and future prospective. Biosci. Rep. 2015, 35:e00191.
100. Windrum P., Morris T.C., Drake M.B., Niederwieser D., Ruutu T., Subcommittee E.C.L.W.P.C. Variation in dimethyl sulfoxide use in stem cell transplantation: a survey of EBMT centres. Bone Marrow Transplant. 2005, 36:601-603.
101. Thirumala S., Goebel W.S., Woods E.J. Clinical grade adult stem cell banking. Organogenesis 2009, 5:143-154.
102. Thirumala S., Zvonic S., Floyd E., Gimble J.M., Devireddy R.V. Effect of various freezing parameters on the immediate post-thaw membrane integrity of adipose tissue derived adult stem cells. Biotechnol. Prog. 2005, 21:1511-1524.