早胜牛胚胎和耳缘成纤维细胞分离培养与鉴定

为了建立早胜牛胚胎和耳缘成纤维细胞体外分离培养体系,采用植块法和酶消化法制备这两种组织成纤维细胞。结果显示,用植块法制备了耳缘成纤维细胞,组织块直径较小的成活率较高且生长速度较快;用酶消化法制备了胚胎和耳缘两种组织成纤维细胞,复苏后细胞的存活率:胚胎(96.8%)＞耳(94.7%)。2种组织成纤维细胞生长曲线均呈"S"型,胚胎组织(倍增时间29.7 h)生长速度快于耳组织(倍增时间31.2 h)。用免疫组织化学染色鉴定证实,分离培养的成纤维细胞中间丝被染成棕黄色,波形蛋白呈阳性反应。

Isolation and Identification of Embryo and Ear Marginal Fibroblasts in Zaosheng Cattle

The embryo and ear marginal fibroblasts in Zaosheng cattle were isolated and identified in vitro using enzyme digestion and explant culture.The survival rate of ear marginal fibroblasts cultured with smaller explant were higher than that with bigger tissue.The growth curves of the embryo and ear marginal fibroblasts cultured with enzyme digestion were sigmoid type.The doubling time of embryo fibroblasts(29.7 h) was shorter than ear marginal bibroblast(31.2 h).The survival rate of defrozen embryo fibroblasts(96.8%) was higher than that of ear marginal fibroblasts(94.7%).The immunohistochemical staining was used to identify that intermediate filament proteins of isolated fibroblasts were stained tan,showing vimentin reaction is positive.