Effect of perfusion flow rate on proliferation and osteoblastic differentiation of human mesenchymal stem cells
- VernacularTitle:灌注型生物反应器中流速对人骨髓间充质干细胞增殖及成骨分化的影响
- Author:
Xiao-jiang SUN
1
;
Ke-rong DAI
1
;
You-zhuan XIE
1
;
Ting-ting TANG
1
;
Zhi-feng YU
1
;
Shu-hong ZHANG
1
Author Information
1. Shanghai Key Laboratory of Orthopeadic Implants, Department of Orthopaedics, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine
- Publication Type:Journal Article
- Keywords:
Bioreactor;
Flow shear stress(FSS);
Human mesenchymal stem cells (hMSCs);
Proliferation;
Osteoblastic differentiation
- From:
Journal of Medical Biomechanics
2012;27(5):E582-E587
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of different perfusion flow rates on proliferation and osteoblastic differentiation of human mesenchymal stem cells (hMSCs) in large scale β-TCP (tricalcium phosphate) scaffold at perfusion bioreactor. Methods hMSCs isolated from iliac bone marrow aspiration were loaded into large scale β-TCP scaffold and cultured in perfusion bioreactor at the perfusion flow rate of 3, 6 or 9 mL/min for 15 days. The culture media were collected for D-glucose consumption assay every 3 days. After perfusion culture for 15 days, the cell-scaffold composites were harvested for assessment of cell viability by MTT colorimetric method, SEM observation and osteogenic gene expression by real-time PCR. Results The proliferation of hMSCs assayed by daily glucose consumption showed that at early stage of culture, cells proliferated faster at flow rate of 9 mL/min than at 3 or 6 mL/min (P<0.001); while at late stage of culture, cells proliferated faster at flow rate of 6 mL/min (P<0.05). The cell viability indicated that the cell-scaffold composites at flow rate of 6 mL/min exhibited the most viable cells (P<0.001). SEM indicated that all the macropores of the scaffold at different flow rates were filled with cellular layers. All cellular layers at flow rate of 3 mL/min were incompact, but that at 9 mL/min were compact; at flow rate of 6 mL/min, the cellular layers were either compact or incompact. Real-time PCR revealed that after perfusion culture for 15 days, the mRNA expression of osteobalstic genes including ALP and OP, were enhanced significantly at flow rate of 6 and 9 mL/min as compared to that at 3 mL/min (P<0.01); however, the 9 mL/min group presented the higher OC expression than 3 and 6 mL/min group (P<0.001). Conclusions At early stage of perfusion culture, the proliferation of hMSCs was promoted at flow rate of 9 mL/min, while at late stage, there was more viable cells in scaffolds at flow rate of 6 mL/min. The osteoblastic differentiation of hMSCs was facilitated with the increase of perfusion flow rate, which was attributed to the increased flow shear stress.
