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Characterization of mesenchymal stem cells (MSCs) from human fetal lung: Potential differentiation of germ cells

机译:人类胎儿肺间充质干细胞(MSCs)的表征:生殖细胞的潜在分化

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Pluripotent mesenchymal stem-like cell lines were established from lungs of 3-4 months old aborted fetus. The cells present the high ex vivo expansion potential of MSC, a typical fibroblast-like morphology and proliferate up to 15 passages without displaying clear changes in morphology. Immunological localization and flow cytometry analyses showed that these cells are positive for OCT4, c-Kit, CD11, CD29, CD44, telomerase, CD106, CD105, CD166, and SSEA1, weakly expression or negative for SSEA1, SSEA3, SSEA4, CD34, CD105 and CD106. These cells can give rise to the adipogenic as evidenced by accumulation of lipid-rich vacuoles within cells identified by Oil-red O when they were induced with 0.5 mM isobutylmethylxanthine, 200 mu M indomethacin, 10 super(-6) M dexamethasone, and 10 mu g/ml of insulin in high-glucose DMEM. Osteogenic lineage cells were generated in 0.1 mu M dexamethasone, 50 mu g/ml ascorbic acid, 10 mM beta -glycerophosphate, which are shaped as the osteoblastic morphology, expression of alkaline phosphatase (AP), and the formation of a mineralized extracellular matrix identified by Alizarin Red staining. Neural cells are observed when the cultures were induced with 2-mercapometal, which are positive for nestin, NF-100, MBP and GFAP. Additionally, embryoid bodies (EBs) and sperm like cells are obtained in vitro differentiation of these lung MSCs induced with 10 super(-5) M retinoic acid (RA). These results demonstrated that these MSCs are pluripotent and may provide an in vitro model to study germ-cell formation and also as a potential source of sperms for male infertility.
机译:从3-4个月大流产胎儿的肺中建立了多能间充质干样细胞系。这些细胞具有高度的MSC体外扩增潜力,这是一种典型的成纤维细胞样形态,最多可增殖15代,但形态没有明显变化。免疫定位和流式细胞仪分析显示,这些细胞对OCT4,c-Kit,CD11,CD29,CD44,端粒酶,CD106,CD105,CD166和SSEA1呈阳性,对SSEA1,SSEA3,SSEA4,CD34,CD105呈弱表达或呈阴性和CD106。这些细胞可引起脂肪形成,如当用0.5 mM异丁基甲基黄嘌呤,200μM吲哚美辛,10个super(-6)M地塞米松和10 mM诱导时,由油红O识别的富含脂质的液泡在细胞中的积累可证明这一点。高糖DMEM中的胰岛素微克/毫升。成骨谱系细胞产生于0.1μM地塞米松,50μg/ ml抗坏血酸,10 mMβ-甘油磷酸酯中,其形成为成骨细胞形态,碱性磷酸酶(AP)的表达以及矿化的细胞外基质的形成通过茜素红染色。当用2-巯基金属诱导培养物时观察到神经细胞,2-巯基金属对巢蛋白,NF-100,MBP和GFAP呈阳性。此外,在体外分化中,用10种super(-5)M维甲酸(RA)诱导了这些肺间充质干细胞,获得了胚状体(EBs)和精子样细胞。这些结果表明,这些MSC具有多能性,可以为研究生殖细胞的形成提供体外模型,也可以作为男性不育症的潜在精子来源。

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