| 摘要: |
| 取材于妊娠终止胚胎生殖腺或生殖嵴及其周围组织,用DMEM+NCS培养液体外培养,分离由人PGCs转化成的类ES细胞,并将其与同源胚胎成纤维细胞一起用胰蛋白酶+ EDTA的无钙镁PBS液消化传代。在原代培养12 h观察到胞体较大、边缘不整、贴壁分裂增殖的PGCs样细胞, 48 h后观察到26处紧密排列呈鸟巢状的类ES细胞集落及集落团。第6 d传代成功,第16 d传至第4代。结果表明,体外培养附植后胚胎能够建成人的类ES细胞系。 |
| 关键词: 原始生殖细胞，胚胎干细胞，分离，传代，克隆，体外培养，人 |
| DOI： |
| 分类号:Q492, |
| 基金项目:国家自然科学基金资助项目（39470663） |
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| Embryonic Stem Cell-like Colonies from Primordial Germ Cells in Human |
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| Abstract: |
| Aborted human fetuses were collected to isolate PGCs-derived ES-like cells from genital gland/ridge and the tissue around,with DMEM+NCS(newborn calf serum) as culture medium.ES-like cells were disaggregated and passaged to gether with homologous embryonic fibroblasts using trypsin+EDTA in Ca2+ and Mg2+ free PBS.At 12 h of primary culture PGCs attaching to the bottom of culture dish with large size and blebbling edge were observed.After 48 h arose 26 discrete and mixed ES-like colonies in which cells were densely packed making the colonies nest-like.On day 6 some colonies succeeded in the first passaging,and subcultured to passage 4 on day 16.The result suggested that ES-like cell line in human could be established by culturing post-implantation embryo. |
| Key words: primordial germ cell,embryonic stem cell,isolation,passage,colony,in vitro culture,human |
