Construction of a general albumin promoter reporter system for real-time monitoring of the differentiation status of functional hepatocytes from stem cells in mouse, rat and human
| Autor: | Xiujuan Wu, Ji Bao, Qiong Wu, Hong Bu, Jing Tang, Mingjun Xie, Lihua Zhu, Yujun Shi, Yujia Wang, Yi Li |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Hepatocyte differentiation hepatocyte differentiation Reporter gene Cell type General Neuroscience Mesenchymal stem cell gene reporter Articles General Medicine Transfection Biology albumin promoter Molecular biology General Biochemistry Genetics and Molecular Biology Viral vector 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Cell culture adenoviral vector 030220 oncology & carcinogenesis stems cells General Pharmacology Toxicology and Pharmaceutics Stem cell |
| Zdroj: | Biomedical Reports |
| ISSN: | 2049-9442 2049-9434 |
| DOI: | 10.3892/br.2017.905 |
| Popis: | Genetic constructs with promoters fused to reporter genes for simultaneous monitoring of cellular events have been the focus of attention in recent years. Adenoviral vectors, which have distinctive characteristics, have been used to monitor the differentiation of stem cells in vitro. In the present study, a modified adenoviral vector was constructed, containing a mouse, rat, and human general albumin promoter sequence fused to a ZsGreen reporter gene, and evaluated its efficiency in different cell types. Two hepatocyte cell lines (Hepa1-6 and HepG2), rat primary hepatocytes, rat bone marrow mesenchymal stem cells (BM-MSCs) and rat BM-MSCs-derived hepatocyte-like cells were transduced with this vector, and the transfection efficiency and functional capabilities of the promoter were evaluated by fluorescent microscopy. The results demonstrated efficient expression of ZsGreen in Hepa1-6 cells, HepG2 cells, rat primary hepatocytes, and rat BM-MSCs-derived hepatocyte-like cells, but not in rat BM-MSCs. In conclusion, the current study demonstrates a simple, high-efficiency, general tool for real-time monitoring of the differentiation status of hepatocytes from stem cells in mice, rats, and humans. This tool may be useful for evaluating different protocols to generate functional hepatocytes from stem cells in multiple species. |
| Databáze: | OpenAIRE |
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