Optimization of an effective directed differentiation medium for differentiating mouse bone marrow mesenchymal stem cells into hepatocytes in vitro.
We have used a uniform design to explore the most effective directed differentiation medium (MEDDM) for differentiating mouse bone marrow mesenchymal stem cells (mMSCs) into hepatocytes. Our methods involved arranging eight differentiation medium groups following uniform design. Flow cytometry was used to evaluate the percentage of ALB+ and CK18+ cells in each group. Factors and their concentrations in the MEDDMs were then identified. The MEDDMs were evaluated by their ability to differentiate mMSCs into hepatocytes by RNA and protein expressions and synthesis functions. FGF at 35 ng/ml and OSM at 30 ng/ml in the medium yielded the highest percentage of ALB+ and CK18+ cells. During directed differentiation using MEDDMs, ALB, CK18, TTR, AFP mRNAs were expressed. ALB and CK18 proteins were detected in the cells. The differentiated cells produced albumin and urea in a time dependent manner. Uniform design was adequate for choosing the MEDDM of mMSCs. MEDDM containing 35 ng/ml FGF and 30 ng/ml OSM was effective in differentiating mMSCs into hepatocytes.
Shi
Xiao-Lei
XL
Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, Medical College of Nanjing University, No. 321 Zhongshan Road, Nanjing 210008, PR China.
Mao
Liang
L
Xu
Bi-Yun
BY
Xie
Ting
T
Zhu
Zhang-Hua
ZH
Chen
Jun-Hao
JH
Li
Lei
L
Ding
Yi-Tao
YT
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D003470:Culture Media; D046508:Culture Techniques; D004815:Epidermal Growth Factor / Q000378:metabolism; D051519:Fibroblast Growth Factor 4 / Q000378:metabolism; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D053683:Oncostatin M / Q000378:metabolism; D010641:Phenotype
D015415:Biomarkers; D003470:Culture Media; D051519:Fibroblast Growth Factor 4; D053683:Oncostatin M; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor