原始生殖细胞的人类胚胎干细胞克隆

Embryonic Stem Cell like Colonies from Primordial Germ Cells in Human

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１２ｈ观察到胞体较大、边缘不整、贴壁分裂增殖的ＰＧＣｓ样细胞，４８ｈ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６ｄ传代成功，第１６ｄ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。

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 Ca 2+ and Mg 2+ 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.