Expansion of human mesenchymal stem cells in a fixed-bed bioreactor system based on non-porous glass carrier--part A: inoculation, cultivation, and cell harvest procedures.
Human mesenchymal stem cells (hMSC) are a promising cell source for several applications of regenerative medicine. The cells employed are either autologous or allogenic; by using stem cell lines in particular, allogenic cells enable the production of therapeutic cell implants or tissue engineered implants in stock. For these purposes, the generally small initial cell number has to be increased; this requires the use of bioreactors, which offer controlled expansion of the hMSC under GMP-conform conditions. In this study, divided into part A and B, a fixed bed bioreactor system based on non-porous borosilicate glass spheres for the expansion of hMSC, demonstrated with the model cell line hMSC-TERT, is introduced. The system offers convenient automation of the inoculation, cultivation, and harvesting procedures. Furthermore, the bioreactor has a simple design which favors its manufacturing as a disposable unit. Part A is focused on the inoculation, cultivation, and harvesting procedures. Cultivations were performed in lab scales up to a bed volume of 300 cm³. The study showed that the fixed bed system, based on 2-mm borosilicate glass spheres, as well as the inoculation, cultivation, and harvesting procedures are suitable for the expansion of hMSC with high yield and vitality.
Weber
Christian
C
Institute of Biopharmaceutical Technology, University of Applied Sciences Giessen-Friedberg, Giessen, Germany.
Freimark
Denise
D
Pörtner
Ralf
R
Pino-Grace
Pablo
P
Pohl
Sebastian
S
Wallrapp
Christine
C
Geigle
Peter
P
Czermak
Peter
P
D057205:Automation, Laboratory; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation*; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004209:Disposable Equipment; D004867:Equipment Design; D005898:Glass; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D010101:Oxygen Consumption; D013499:Surface Properties
D005947:Glucose; D010100:Oxygen