title: Optimization of an effective directed differentiation medium for differentiating mouse bone marrow mesenchymal stem cells into hepatocytes in vitro.
abstract: 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.
authors.affiliation: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, Medical College of Nanjing University, No. 321 Zhongshan Road, Nanjing 210008, PR China.
authors.forename: Xiao-Lei
authors.initials: XL
authors.lastname: Shi
authors.forename: Liang
authors.initials: L
authors.lastname: Mao
authors.forename: Bi-Yun
authors.initials: BY
authors.lastname: Xu
authors.forename: Ting
authors.initials: T
authors.lastname: Xie
authors.forename: Zhang-Hua
authors.initials: ZH
authors.lastname: Zhu
authors.forename: Jun-Hao
authors.initials: JH
authors.lastname: Chen
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Yi-Tao
authors.initials: YT
authors.lastname: Ding
mesh_terms: 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
chemical_list: D015415:Biomarkers; D003470:Culture Media; D051519:Fibroblast Growth Factor 4; D053683:Oncostatin M; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor