延边奶山羊耳成纤维细胞体外培养的研究

Study of ear fibroblasts in vitro culture of Yanbian dairy goat

为了给延边奶山羊转基因核移植及乳腺生物反应器研究提供足够且状态良好的细胞,采取延边奶山羊耳部组织块,利用组织块贴壁法进行原代培养和传代培养,对于纯化了的成纤维细胞观察其生长状态,并对4代以后的成纤维进行生长曲线、核型分析,并对细胞进行冷冻复苏试验,检测细胞活力。结果表明,4～13代体外培养成纤维细胞的细胞形态及核型基本正常,13代以后,细胞染色体变异比例增加,15代以后,细胞生长速度减慢;冷冻复苏后的细胞除生长速度减慢外,其它细胞生物学特征均正常,基本符合体细胞克隆、转基因等试验的基本要求,可以用于试验研究。

In order to provide the more cells of good condition for the transplantation of transgenic and mammary gland bioreactor in Yanbian dairy goat, took the ear tissue blocks of Yanbian dairy goat, cul-tured primarily and subculture with tissue block adhering wall method. Observed the growth state of puri-fied fibroblast, and maked cell growth curve, karyotype analysis, and thaw test of fibroblasts after four generations, cell viability was detected. The results showed that cell morphology and karyotype of 4-13 generations fibroblasts was normal in vitro cultured, but after the 13th generation, the proportion of chro-mosomal aberration increased, after the 15th generation, cell growth slowed down; the cells after cryopr-eservation, in addition to slow down the growth rate, other biological characteristics were normal, in line with the basic requirements of somatic cell cloning and the transgenic, can be used in the research.