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Human amniotic epithelial cells differentiate into cells expressing germ cell specific markers when cultured in medium containing serum substitute supplement

机译:在含有血清替代品补充剂的培养基中培养时,人羊膜上皮细胞分化为表达生殖细胞特异性标志物的细胞

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Background Human amniotic epithelial cells (hAECs) maintain the plasticity of pregastrulation embryonic cells, having the potential to differentiate into all three germ layers. The potential of these cells to differentiate into cells expressing germ cell specific markers has never been described before. Methods In the present study, hAECs were cultured in medium containing serum substitute supplement (SSS). Gene and protein expression of germ cell and oocyte specific markers was assessed by reverse transcription-polymerase chain reaction ( RT-PCR), immunofluorescence staining and flow activated cell sorter analysis (FACS) in hAECs at different time points during the differentiation into cells expressing germ cell specific markers. Results When cultured with SSS, already at passage 1, hAECs start to express the germ cell specific genes C-KIT, DAZL, VASA and ZP3 and at passage 5 large round cells, resembling oocytes, appeared. The cells express the germ cell specific marker DAZL, the oocyte specific markers GDF9 and ZP3 and the meiosis specific markers DMC1 and SCP3 at the protein level. Conclusions From our preliminary results we can conclude that hAECs have the potential to differentiate into cells expressing germ cell specific markers.
机译:背景技术人羊膜上皮细胞(hAEC)维持妊娠前胚胎细胞的可塑性,具有分化为所有三个胚层的潜力。这些细胞分化成表达生殖细胞特异性标志物的细胞的潜力从未被描述过。方法在本研究中,将hAECs培养在含有血清替代品补充剂(SSS)的培养基中。通过逆转录-聚合酶链反应(RT-PCR),免疫荧光染色和流式细胞分选分析(FACS)在hAEC中的不同时间点评估生殖细胞和卵母细胞特异性标志物的基因和蛋白质表达细胞特异性标记。结果当用SSS培养时,已经在第1代中,hAEC开始表达生殖细胞特异性基因C-KIT,DAZL,VASA和ZP3,并在第5代出现了类似卵母细胞的大圆形细胞。这些细胞在蛋白质水平上表达生殖细胞特异性标记DAZL,卵母细胞特异性标记GDF9和ZP3以及减数分裂特异性标记DMC1和SCP3。结论从我们的初步结果中,我们可以得出结论,hAEC具有分化成表达生殖细胞特异性标记物的细胞的潜力。

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