利用动物耳皮肤组织培养成纤维细胞的研究

对采集的动物耳部皮肤样品采用将组织块剪碎后直接贴附于细胞培养瓶底部培养的方法与酶消化法对动物体细胞进行原代培养 ,前者使得对不同品种绵羊、山羊、牛、北山羊的原代体细胞出现率及可传代率均达到 1 0 0 %。两种培养方法在成纤维细胞出现及可进行传代时间上有所差别。对于在体细胞传代培养中出现的上皮样细胞与成纤维样细胞混合生长的问题根据两者贴壁紧实程度的不同 ,用 0 .0 5%的胰酶—EDTA对其进行不同时间的消化 ,将两种不同类型的细胞进行分离和纯化。纯化的体细胞经数次传代培养后进行冷冻—解冻后仍具有正常的传代能力。对不同代次绵羊和牛的体细胞进行的核型分析表明 ,在体外培养至 2 0～ 30代 (绵羊胎儿细胞培养至 45代 )成纤维细胞的染色体倍性及数目未发现异常

Sdudy on in vitro culture of fibroblast cells using animal ear culture

Adult animals ear skin cells were culture and passaged using methods of cut in to pieces directly and enzyme digest. The former method had a 100% of primary passage y fibroblast cells present rate and successful passage rate for all the experiment animals including sheep, goat, cow ,and north goat. Two methods had a differences between primary fibroblast cells present rate and time of can be passaged. Using different digested time to purified fibroblast cells with epithelial cells. Purified fibroblast cells still have normal capacity of passage after freeze-thawed. Karyotype analyze indicate that after 20-30 passage of in vitro culture(sheep fetus cells cultured up to 45 passage) no abnormal of chromosomes' number and ploidy of fibroblast cells be found.