| 摘要: |
| 目的探讨NKX6.1对PDX1诱导人胎肝间充质干细胞(fetal liver-derived mesenchymal stem cells,FL-MSCs)向胰岛B细胞分化的协同作用及可能机制，以获取足够用于移植的胰岛B样细胞。方法构建含PDX1与NKX6.1双基因的重组腺病毒载体，用该载体感染FL-MSCs并联合相应的细胞因子分步诱导,检测诱导后的细胞中PDX1和NKX6.1基因以及NGN3、NeuroD1/Beta2、MafA因子和胰岛素等多种胰腺B细胞相关分子的表达情况。结果腺病毒载体转染24 h后FL-MSCs细胞即开始表达PDX1与NKX6.1基因，检测显示诱导后的细胞先后开始表达NGN3、NeuroD1和MafA等因子，持续稳定表达胰岛素等B细胞相关分子；且这些分子表达存在时序性。结论PDX1与NKX6.1联合细胞因子在体外能有效诱导FL-MSCs分化为胰岛B样细胞；可能是通过先后活化的NGN3、NeuroD1和MafA等转录因子，限定细胞向内分泌前体细胞、进一步向B内分泌前体细胞、B细胞分化发育。 |
| 关键词: 胰腺十二指肠同源框1 NKX6.1 胎肝 间充质干细胞 腺病毒载体 |
| DOI：10.3724/SP.J.1008.2010.0258 |
| 投稿时间：2009-08-01修订日期：2009-12-28 |
| 基金项目:安徽省教育厅自然科学基金(2006kj303B). |
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| NKX6.1 combined with PDX1 induces mesenchymal stem cell differentiation into B-like cells |
| TANG Xiao-long1,2*, ZHANG Rong-bo2, WANG Xue-feng2 |
| (1. Clinical Laboratory, Guangdong Provincial Hospital of TCM, Guangzhou 510120, Guangdong, China;2. Institute of Biomedical Engineering, Medical College, Anhui University of Science and Technology, Huainan 232001, Anhui, China) |
| Abstract: |
| ObjectiveTo study the synergistic effect of NKX6.1 and PDX1 in inducing differentiation of fetal liver-derived mesenchymal stem cells(FL-MSCs) into the pancreatic B cells and to explore the underlying mechanisms, so as to obtain enough islet-like body for transplantation. MethodsRecombinant adenovirus vector harboring both PDX1 and NKX6.1 genes was constructed, and the vector was used to infect FL-MSCs. Then a series of cytokines were used to induce the differentiation of infected FL-MSCs into pancreatic B cells. The expressions of PDX1, NKX6.1 gene, transcription factors NGN3, NeuroD1/Beta2, MafA as well as C-peptide were examined. ResultsPDX1 and NKX6.1 were detected in FL-MSCs cells 24 h after infection; cells began to express NGN3, NeuroD1, and MafA and stably expressed pancreatic B cell related factors including insulin after induction.The expression of these molecules was in a certain order. ConclusionPDX1, NKX6.1 combined with a series of cytokines can effectively induce FL-MSCs to differentiate into pancreatic islet B cells in vitro, which might be through activation of transcription factors NGN3, NeuroD1, and MafA in turn, inducing FL-MSCs to differentiate towards endocrine precursor cells,B endocrine precursor cells and B cells in turn. |
| Key words: pancreatic duodenal homeobox 1 NKX6.1 fetal liver mesenchymal stem cells the adenovirus vector |
