[Construction and characteristics analysis of somatic cell lines from common carp Cyprinus carpio].

Shi yan sheng wu xue bao Pub Date : 2005-02-01
Fan Hua Meng, Hong Bin Yin, Zhong Wu Sun, Wei Duo Chen
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Abstract

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 characteristics 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 degrees C; (3) there isn't any change in cell chromosomes, which still are normal diploid. However, with, the generations increasing, 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 degrees C. 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.

鲤鱼(Cyprinus carpio)体细胞的构建及特性分析。
取鲤鱼喙侧和鱼鳍的上皮细胞进行体外培养。结果,尾鳍细胞成功地代代相传,并形成了细胞系。在培养过程中,观察传代细胞并分析其生物学特性,包括形态分析、生长曲线绘制、无性系繁殖率检测、温度适应性观察、染色体计算和冷冻细胞解冻研究。根据观察结果,我们发现(1)成纤维细胞有增加的趋势。鲤鱼嘴侧和尾鳍上皮是培养细胞的最佳材料;(2)最适培养温度为27℃;(3)细胞染色体没有变化,仍是正常的二倍体。然而,随着世代的增加,染色体模式在正常范围内有下降的趋势。为了保证细胞核移植后的遗传稳定性和存活率,我们推断4-10代的供体细胞是核移植实验的最佳选择。在27℃条件下培养48 ~ 72h,细胞生长良好。建议在使用DMSO保存细胞时,温度应逐渐降低。到目前为止,细胞已经传递了30代,部分细胞被保存在液氮中。
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