Human bone marrow mesenchymal stem cells can express insulin and key transcription factors of the endocrine pancreas developmental pathway upon genetic and/or microenvironmental manipulation in vitro
Human bone marrow mesenchymal stem cells can express insulin and key transcription factors of the endocrine pancreas developmental pathway upon genetic and/or microenvironmental manipulation in vitro
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DOI：
10.1634/stemcells.2004-0123
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发表时间：
2005-04-01
期刊：
影响因子：
5.2
通讯作者：
Benhamou, PY
中科院分区：
文献类型：
--
作者：
Moriscot, C;De Fraipont, F;Benhamou, PY
多能干细胞可以通过塑料粘附从骨髓中选择、扩增和培养。它们不仅能够分化成多种细胞类型，包括软骨、骨、脂肪和纤维组织以及骨髓支持基质，而且能够分化成中胚层（内皮）、神经外胚层或内胚层（肝细胞）谱系。我们的目标是表征人间充质干细胞（hMSCs）的多潜能能力，并评估其体外分化为胰岛素分泌细胞的能力。从健康供体中获得hMSCs，通过塑料粘附进行选择，并在用编码小鼠IPF 1、HLXB 9和FOXA 2转录因子的腺病毒感染之前和之后通过荧光激活细胞分选仪和逆转录聚合酶链反应分析进行表型分析，所述转录因子参与内分泌发育途径的早期。我们发现天然hMSC具有多能表型（OCT 4表达和高端粒长度），并以低水平组成性表达NKX 6 -1，但缺乏与β细胞分化有关的所有其他转录因子。在所有的hMSCs中，我们检测到细胞角蛋白18和19的mRNA，上皮标记物存在于胰腺导管细胞，而前转化酶1/3 mRNA的表达仅在一些hMSCs中检测到。有或没有胰岛共培养或胰岛条件培养基的情况下，IPF 1、HLXB 9和FOXA 2的异位表达导致胰岛素基因表达。总之，我们的研究结果表明，在体外培养的人骨髓干细胞能够分化成胰岛素表达细胞的机制，涉及几个转录因子的β细胞发育途径时，在适当的微环境中培养。
Multipotential stem cells can be selected from the bone marrow by plastic adhesion, expanded,and cultured. They are able to differentiate not only into multiple cell types, including cartilage, bone, adipose and fibrous tissues, and myelosupportive stroma, but also into mesodermal (endothelium), neuroectodermal, or endodermal (hepatocytes) lineages. Our goal was to characterize the multipotential capacities of human mesenchymal stem cells (hMSCs) and to evaluate their ability to differentiate into insulin-secreting cells in vitro. hMSCs were obtained from healthy donors, selected by plastic adhesion, and phenotyped by fluorescence-activated cell sorter and reverse transcription-polymerase chain reaction analysis before and after infection with adenoviruses coding for mouse IPF1, HLXB9, and FOXA2 transcription factors involved early in the endocrine developmental pathway. We found that native hMSCs have a pluripotent phenotype (OCT4 expression and high telomere length) and constitutively express NKX6-1 at a low level but lack all other transcription factors implicated in beta-cell differentiation. In all hMSCs, we detected mRNA of cytokeratin 18 and 19, epithelial markers present in pancreatic ductal cells, whereas proconvertase 1/3 mRNA expression was detected only in some hMSCs. Ectopic expression of IPF1, HLXB9, and FOXA2 with or without islet coculture or islet-conditioned medium results in insulin gene expression. In conclusion, our results demonstrated that in vitro human bone marrow stem cells are able to differentiate into insulin-expressing cells by a mechanism involving several transcription factors of the beta-cell developmental pathway when cultured in an appropriate microenvironment.