小鼠外胎盘锥次级滋养层巨细胞的分离、培养与鉴定

目的：建立小鼠外胎盘锥次级滋养层巨细胞的分离、培养方法.方法：在小鼠妊娠第8 d,通过机械分离和组织块翻转干涸法分离、培养小鼠外胎盘锥次级滋养层巨细胞;免疫细胞化学鉴定细胞来源与纯度;酶谱方法检测其所分泌基质金属蛋白酶2和9（matrix metalloproteinase-2,9,MMP-2,9）结果：利用机械分离法分离外胎盘锥可生长于Matrigel包被的培养板上,并生长出次级滋养层巨细胞,表达特异性标志物Cytokeratin;体外培养24 h、48 h后,外胎盘锥滋养层巨细胞的粘附率分别为（83.3±9.1）%和（92.4±7.3）%;扩展率分别为（45.5±5.3）%和（56.4±6.8）%;及其所分泌的MMP-2,9的24 h光密度值分别为（87±4.7）和（351.5±25.2）;48 h分别为（186±40.2）和（556.5±61.5）.结论：采用机械分离和组织块翻转干涸法,可简单、快捷地获得高纯度小鼠外胎盘锥次级滋养层巨细胞.

Isolation,Culture and Identification of Second Trophoblast Giant Cells of Mouse Ecoplacental Cones

To establish the method of isolation and culture of second trophoblast giant cells （sTGC） of mouse ecoplacental cone. Methods： Ectoplacental cones were isolated and cultured by mechanical separation and methods of tissue turnover and dry on day 8 of mouse pregnancy. Immunocytoehemistry revealed the cell characteristics and purity ot sTGC. Secreted MMP-2, 9 were detected by gelatin zymography. Results： Ectoplacental cone separated by mechanical method can be grown in Matrigel-coated culture plate, and sTGC could be grown out of EPC and exhibited positive staining for cytokeratin. At 24 h and 48 h after initiation of in vitro culture, adhesion rates of ectoplacental cone were （ 83.3 ± 9.1 ） % and （ 92.4 ± 7.3 ） % ; outgrowth rates were （45.5 ± 5.3 ） % and （ 56.4 ± 6.8 ） %. Density values of ectoplacental cone secreted MMP-2 and9 were （87 ±4.7） and （51.5 ±25.2） at 24 h; （186 ±40.2） and （556.5 ± 61.5） at 48, respectively. Conclusion： High purity, sTGC of mouse ecoplacental cone were successfully obtained by methods of mechanical separation and tissue turnover and dry.