| 谢佩怡,胡晓俊,陈俊伟,孟晓春,朱康顺,单鸿.过表达肝细胞核因子4 alpha对人骨髓间充质干细胞向肝样细胞分化的作用[J].中国医学影像技术,2014,30(7):991~995 |
| 过表达肝细胞核因子4 alpha对人骨髓间充质干细胞向肝样细胞分化的作用 |
| Hepatocyte-like cells from human bone marrow mesenchymal stem cells：Effects of hepatocyte nuclear factor 4 alpha gene overexpression |
| 投稿时间：2014-02-19 修订日期：2014-06-05 |
| DOI： |
| 中文关键词: 肝细胞核因子4α 间质干细胞 细胞分化 |
| 英文关键词:Hepatocyte nuclear factor 4 alpha Mesenchymal stem cells Cell differentiation |
| 基金项目:国家自然科学基金（81071206、81172193、81201090、81070349、81371655）、NSFC-广东联合基金重点项目（U1032002）、卫生部临床学科重点项目（164）。 |
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| 中文摘要: |
| 目的 应用转基因技术将肝细胞核因子4 alpha（HNF-4α）转导入人骨髓间充质干细胞（MSCs）内，使其连续过表达HNF-4α并促进MSCs向肝样细胞分化。方法 HNF-4α基因通过慢病毒表达载体pLV/Final-puro-hHNF4α-hrGFP转入人MSCs（UE7T-13细胞）内，用流式细胞分析法检测及分选后，收集hrGFP阳性的UE7T-13细胞进行扩增。体外成肝诱导一周后，免疫荧光染色检测过表达HNF-4α基因的UE7T-13细胞（E7-hHNF-4α细胞）的白蛋白（ALB）和细胞角蛋白（CK18）的表达情况，糖原染色检测UE7T-13细胞及E7-hHNF-4α细胞的糖原储存功能。结果 建立稳定、过表达hHNF-4α基因的E7-hHNF-4α细胞，持续过表达HNF-4α促进MSCs向肝样细胞分化，经过7天体外成肝诱导，E7-hHNF-4α细胞具有成熟肝样细胞表达ALB和CK18蛋白及糖原储存功能。结论 利用Gateway 技术可将HNF-4α高效转导入人MSCs细胞中，并有效促进MSCs向肝样细胞分化。 |
| 英文摘要: |
| Objective To transduce hepatocyte nuclear factor 4 alpha (HNF-4α) by lentiviral vector into human bone marrow mesenchymal stem cells (BM-MSCs) to improve the hepatic differentiation. Methods HNF-4α genes were transduced into UE7T-13 BM-MSCs by lentiviral vector (pLV/Final-puro-hHNF4α-hrGFP). After twice transduction, HNF-4α-transduced MSCs (E7-hHNF-4α cell) expanded in vitro and prepared for fluorescence-activated cell sorting (FACS) analysis. Maturated hepatic functions such as albumin (ALB) and cytokeratin 18 (CK18) of E7-hHNF-4α cells were detected by immunofluorescence staining one week after hepatic differentiated in vitro. Glycogen storage was measured by periodic acid-Schiff (PAS) staining. Results Stable, high-level expression of HNF-4α of UE7T-13 cells were established after infection with HNF-4α lentivirus. The hepatic differentiation from human BM-MSCs were promoted efficiently by HNF-4α. Upon in vitro hepatic culture systems, the differentiated cells showed ALB and CK18 expression and glycogen storage function. Conclusion Gateway technology is helpful for the transduction of HNF-4αinto human MSCs cells, and promoting the cell differentiation of MSCs to hepatocyte-like cells effectively. |
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