蛋鸡子宫内膜细胞的分离培养与鉴定及性腺激素对其增殖的影响

Culture and identification of endometrial cells of laying hen and effects of sex hormones on the proliferation

为探讨蛋鸡子宫内膜细胞的形态学特点、生长规律,本试验采用Ⅰ型胶原酶消化的方法获取蛋鸡子宫内膜上皮细胞和基质细胞,采用免疫组织化学法进行细胞鉴定,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐比色(MTT)法测得细胞生长曲线,并研究不同浓度水平的性腺激素对子宫内膜上皮细胞和基质细胞体外增殖的影响。结果显示,该方法培养的上皮细胞和基质细胞分别对角蛋白质ck18单克隆抗体和波形蛋白质抗体呈阳性;蛋鸡子宫内膜上皮细胞和基质细胞在接种后第2 d开始进入对数生长期,第7 d进入平台期;孕酮(100 nmol/L)能显著促进子宫内膜上皮细胞和基质细胞体外增殖;孕酮(100 nmol/L)和10 nmol/L、100 nmol/L雌激素组合对子宫内膜上皮细胞和基质细胞体外增殖均有显著促进作用。表明本研究成功培养了蛋鸡子宫内膜上皮细胞和基质细胞,并证实这2种细胞的细胞周期均为7 d,一定浓度的孕酮和雌激素及其组合对子宫内膜上皮细胞和基质细胞的体外增殖均有促进作用。

Endometrial epithelial and stromal cells were isolated and identified by type I collagenase digestion and immunohistochemistry, respectively. The growth curves were constructed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tet-razolium bromide ( MTT) assay. The effects of different concentrations of sex hormones, estrogen ( E2 ) and progesterone ( P4 ) , on the proliferation of the endometrial epithelial and stromal cells in vitro was studied. The epithelial and stromal cells were tested positive to keratin ck18 and vimentin antibody. The growth rates of chicken endometrial epithelial and stro-mal cells began to increase logarithmically on day 2, reaching the plateau phase on day 7. The endometrial cells and stromal cells treated with 100 nmol/L P4 proliferated at a significantly higher rate than the control. However, the endometrial epithelial cells and stromal cells treated with 100 nmol/L or 10 nmol/L E2 plus 100 nmol/L P4 proliferated at a significantly higher rate than the control.