摘要
动物细胞的培养技术是1907年哈里逊[1]在淋巴块中对蛙的神经板培养成功开始的,其后近一个世纪以来,陆续成功地培养了哺乳动物、昆虫等各种动物细胞,并广泛用于生物科学的各个分支.鱼类的细胞培养的系统研究和建系实践大约起始于60年代,被公认的真骨鱼类的第一个永久性的细胞系--虹鳟性腺细胞系(RTG-2)是由Wolf[2]建立的.
Epithelia of rostral side and fins from common carp were taken to culture in vitro. As a result, cells of tail fins were successfully passed from generation to generation, and constructed cell lines. During the culture procedure, the passage cells were observed and their biological char- acteristics were analyzed, which consist of morphologic analysis, drawing of growth curve, detection of producing rate for clones, insight of adaptation to temperatures, calculation of chromosomes and investigation for thaw of frozen cells. According to the observation results, we found that (1) there is an increase trend of fibroblast. epithelia of rostral side and tail fin from common carp are the best materials for culturing cells; (2) the optimum culturing temperature is 27℃; (3) there isn't any change in cell chromosomes, which still are normal diploid. However, with the generations in- creasing, the mode of chromosome has an inclination of decrease in normal ranges. We inferred the donor cells from 4-10 generations are the best choice for nuclear transplanted experiments in order to guarantee their genetic stabilities and survival rates after transplanting the nuclei. And, the cells will grow extremely well when cultured for 48-72h at 27℃. We suggest the temperature should be descended little by little involving in preserving cells with DMSO. So far, the cells were passed 30th generations and part of them was preserved into liquid nitrogen.
出处
《实验生物学报》
CSCD
北大核心
2005年第1期80-84,共5页
Acta Biologiae Experimentalis Sinica
基金
中国水产科学研究院基金(2001-3-1)黑龙江省基金"鱼类克隆技术的研究"
