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Rat bone marrow derived mesenchymal progenitor cells support mouse ES cell growth and germ-like cell differentiation.

机译:大鼠骨髓来源的间充质祖细胞支持小鼠ES细胞的生长和细菌样细胞的分化。

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Mouse embryonic fibroblasts (MEFs) have been used as feeder cells to support the growth of mouse embryonic stem cell (mESC) and primordial germ cells (PGC) in culture for many years. However, MEF preparation is a complex and tedious task. Recently, there are reports indicating that the microenvironment provided by bone marrow stromal cells could support the survival of embryonic-like stem cells in bone marrow. In this report, rat bone marrow derived mesenchymal progenitor cells (MPC) were used as feeder cells to culture mouse Oct4-GFP ES cell and ES cell derived germ cells. FACS results show that similar to MEF, rat MPC could efficiently support growth of the mouse Oct4-GFP ES cell line in culture (MPC 85.5 +/- 5.1% vs MEF 84.1 +/- 6.2%). ES cells could be subcultured for >15 passages without losing morphological characteristics. The cultured cells expressed stem cell marker alkaline phosphatase, Oct4, Sox2, and SSEA-1. Furthermore, rat MPC cells were able to support survival of germ cells isolated from mouse Oct4-GFP ES cell formed embryoid bodies (EB). After induction by retinoic acid for 7 days, some isolated cells differentiated to spermatogonial stem-like cells, expressing Mvh, Stra-8, Hsp90-a, integrinb1 and a6. Compared with traditional MEF culture systems, the rat MPC culture system is effective in supporting ES cell growth and is easy to prepare.
机译:小鼠胚胎成纤维细胞(MEF)已用作饲养细胞,以支持培养中的小鼠胚胎干细胞(mESC)和原始生殖细胞(PGC)的生长。但是,MEF的准备是一项复杂而繁琐的任务。最近,有报道表明,骨髓基质细胞提供的微环境可以支持胚胎样干细胞在骨髓中的存活。在此报告中,大鼠骨髓来源的间充质祖细胞(MPC)被用作饲养小鼠Oct4-GFP ES细胞和ES细胞来源的生殖细胞的饲养细胞。 FACS结果表明,与MEF相似,大鼠MPC可以有效地支持培养的小鼠Oct4-GFP ES细胞系的生长(MPC 85.5 +/- 5.1%与MEF 84.1 +/- 6.2%)。 ES细胞可以传代培养> 15代,而不会失去形态特征。培养的细胞表达干细胞标记碱性磷酸酶,Oct4,Sox2和SSEA-1。此外,大鼠MPC细胞能够支持从小鼠Oct4-GFP ES细胞形成的胚状体(EB)分离的生殖细胞的存活。维甲酸诱导7天后,一些分离的细胞分化为精原干样细胞,表达Mvh,Stra-8,Hsp90-a,整联蛋白a1和a6。与传统的MEF培养系统相比,大鼠MPC培养系统可有效支持ES细胞生长,并且易于制备。

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