[Cultivation of human mesenchymal stem cells on macroporous CultiSpher G microcarriers].
Adult human bone marrow-derived mesenchymal stem cells (MSCs) were cultured on microcarriers in spinner flasks and were compared with those in conventional culture in 12-well plates. For the production of adherently growing MSCs, macroporous CultiSpher G gelatin microcarriers were used in concentration of 1 g/L. The cells were seeded in a density of 5 x 10(4) cells/ml in both spinner culture and conventional stationary culture. The result showed that after 7 days of cultivation a maximum viable cell concentration of 5.15 x 10(5) cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675 x 10(5) cells/ml on day 5 in conventional stationary culture. Lactate was produced up to 12.06 mmol/L in spinner culture and up to 13.10 mmol/L in stationary culture, and glucose was consumed up to 7.38 mmol/L and 5.37 mmol/L respectively. The average lactate yield on glucose consumption in spinner culture was only 1.63, lower than that in stationary culture 2.44. This indicated that the energy metabolism in spinner culture was significantly more efficient than that in conventional culture. After spinner culture for 12 days, the MSCs maintain the characteristics of stem cells. It is concluded that the microcarrier culture system is a suitable way to expand the seeding cells for tissue engineering.
Wu
Qing-Fa
QF
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Wu
Chu-Tse
CT
Dong
Bo
B
Wang
Li-Sheng
LS
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002455:Cell Division; D002470:Cell Survival; D005434:Flow Cytometry; D005947:Glucose / Q000378:metabolism; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D007773:Lactates / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors
D015703:Antigens, CD; D006684:HLA-DR Antigens; D007773:Lactates; D005947:Glucose