CAJ | 학술논문

为探讨适合水牛胎儿成纤维细胞(BFF)的体外培养体系,采用常规组织块法和胰蛋白酶消化法原代培养BFF均获得了较多的成纤维细胞,但后者所得细胞的活力不如前者高,且死细胞也较多;传代或冻存成纤维细胞时用4℃预冷的胰蛋白酶室温下消化所得的细胞比37℃热消化的细胞更圆、更有光泽;跟踪32代的细胞冷冻复苏率均达70%～80%;染色体分析结果显示,二倍体细胞所占比例始终保持在80%～90%之间,各代细胞(5th、10th、15th)之间差异不显著(P>0.05).结果表明,组织块法原代培养、4℃预冷胰蛋白酶室温消化传代细胞的培养体系比较适合水牛胎儿成纤维细胞的培养.
위탐토괄합수우태인성섬유세포(BFF)적체외배양체계,채용상규조직괴법화이단백매소화법원대배양BFF균획득료교다적성섬유세포,단후자소득세포적활력불여전자고,차사세포야교다;전대혹동존성섬유세포시용4℃예랭적이단백매실온하소화소득적세포비37℃열소화적세포경원、경유광택;근종32대적세포냉동복소솔균체70%～80%;염색체분석결과현시,이배체세포소점비례시종보지재80%～90%지간,각대세포(5th、10th、15th)지간차이불현저(P>0.05).결과 표명,조직괴법원대배양、4℃예랭이단백매실온소화전대세포적배양체계비교괄합수우태인성섬유세포적배양.
This paper investigated the culture system of buffalo fetal fibroblasts (BFF) in vitro. A lot of buffalo fetal fibroblasts were obtained with both explant primary culture and digestion primary culture, but cellular vitality was higher in the former more than the latter, and the latter had many dead cells. It was better to digest cells using pre-cold 0.25% trypsin than hot 0.25% trypsin. Moreover, 70%～80% of cells survived after freezing and thawing when subculture 32 passages. Karyotype analysis indicated that 80%～90% of cells had normal karyotypes, which did not show significant difference among the 5th, 10th and 15th passages (P>0.05). The results showed that this culture system was suitable to culture buffalo fetal fibroblasts.