Talaei-Khozani Tahereh, Khodabandeh Zahra, Jaberipour Mansooreh, Hosseini Ahmad, Bahmanpour Soghra, Vojdani Zahra
Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran;
Rom J Morphol Embryol. 2015;56(4):1365-70.
The research on Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) from the umbilical cord suggests promising therapeutic use for hepatocyte replacement therapy. One of the highly conserved members of the nuclear receptor superfamily in the liver is hepatocyte nuclear factor-4α (HNF4α), involved in hepatocyte differentiation. The objectives of this study were to determine the effects of two- and three-dimensional (2D and 3D) cultures of WJ-MSCs on hepatocyte differentiation. MSCs were isolated from human Wharton's jelly, characterized by flow cytometry, and differentiated toward osteogenic and adipogenic lineage. WJ-MSCs were cultured in 2D collagen films and 3D collagen scaffolds in the presence of hepatogenic media with or without pre-treatment with fibroblast growth factor-4 (FGF4) for 21 days. The expression of HNF4α was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). According to flow cytometry data, the cells isolated from Wharton's jelly were shown to express MSC markers. HNF4α expression analysis revealed that pre-exposing the cells with FGF4 was more effective in hepatocyte differentiation. 3D cultures also improve the expression of HNF4α compared with 2D culture system. In conclusion, the combination of FGF4 and 3D culture improved hepatocyte differentiation. It seems 3D interaction of the cells improved the hepatogenesis.
对脐带华通氏胶源性间充质干细胞（WJ-MSCs）的研究表明，其在肝细胞替代治疗方面具有广阔的治疗应用前景。肝细胞核因子-4α（HNF4α）是肝脏中核受体超家族高度保守的成员之一，参与肝细胞分化。本研究的目的是确定WJ-MSCs的二维和三维（2D和3D）培养对肝细胞分化的影响。从人华通氏胶中分离出间充质干细胞，通过流式细胞术进行表征，并向成骨和成脂谱系分化。WJ-MSCs在二维胶原膜和三维胶原支架中培养，在有或没有用成纤维细胞生长因子-4（FGF4）预处理的成肝细胞培养基中培养21天。使用定量实时聚合酶链反应（qRT-PCR）评估HNF4α的表达。根据流式细胞术数据，从华通氏胶中分离出的细胞显示表达间充质干细胞标志物。HNF4α表达分析表明，用FGF4预先处理细胞对肝细胞分化更有效。与二维培养系统相比，三维培养也提高了HNF4α的表达。总之，FGF4和三维培养的组合改善了肝细胞分化。细胞的三维相互作用似乎改善了肝细胞生成。
