Saldeen Johan, Kriz Vitezslav, Agren Nina, Welsh Michael
Department of Medical Cell Biology, Uppsala University, Biomedical Centre, P.O. Box 571, SE-751 23 Uppsala, Sweden.
Biochem Biophys Res Commun. 2006 Jun 2;344(2):517-24. doi: 10.1016/j.bbrc.2006.03.129. Epub 2006 Apr 19.
The potential use of embryonic stem (ES) cells for cell therapy of diabetes requires improved methods for differentiation and isolation of insulin-producing beta-cells. The signal transduction protein SHB may be involved in both angiogenesis and beta-cell development. Here we show that cells expressing the pancreatic endodermal marker PDX-1 appear in the vicinity of vascular structures in ES cell-derived embryoid bodies (EBs) cultured in vitro. Moreover, overexpression of SHB as well as culture of EBs in presence of the angiogenic growth factors PDGF or VEGF enhanced the expression of PDX-1 and/or insulin mRNA. Finally, expression of GFP under control of the PDX-1 promoter in EBs allowed for the enrichment by FACS of cells expressing PDX-1, C-peptide, and insulin as determined by immunofluorescence. It is concluded that SHB and angiogenic factors promote the development of cells expressing PDX-1 and insulin in EBs and that such cells can be separated by FACS.
胚胎干细胞（ES细胞）在糖尿病细胞治疗中的潜在应用需要改进胰岛素分泌β细胞的分化和分离方法。信号转导蛋白SHB可能参与血管生成和β细胞发育过程。在此我们表明，在体外培养的ES细胞来源的胚状体（EBs）中，表达胰腺内胚层标志物PDX-1的细胞出现在血管结构附近。此外，SHB的过表达以及在血管生成生长因子PDGF或VEGF存在下培养EBs可增强PDX-1和/或胰岛素mRNA的表达。最后，在EBs中由PDX-1启动子控制的GFP表达使得通过荧光激活细胞分选（FACS）能够富集经免疫荧光测定表达PDX-1、C肽和胰岛素的细胞。结论是，SHB和血管生成因子促进EBs中表达PDX-1和胰岛素的细胞的发育，并且此类细胞可通过FACS分离。
