革胡子鲶全血培养制备染色体条件探索

Exploration on the Conditions of Whole Blood Cultivation and Chromosome Preparation for Clarias leather

[目的]探究革胡子鲶全血培养及染色体制备的条件方法。[方法]革胡子鲶消毒后用含有肝素的一次性注射器采血,采用RP-MI1640全培养基进行体外全血细胞培养。通过对革胡子鲶血液的细胞培养温度、秋水仙素浓度及加入时间、低渗时间及温度等各种条件的筛选,建立起较成熟的革胡子鲶全血细胞的培养和染色体制备的试验方法。[结果]5 ml培养基中加入0.2 ml全血,26℃下培养72h,在结束培养前6 h加入终浓度0.1μg/ml秋水仙素,低渗35 min,可获得较好的染色体分裂相。[结论]结果为进一步研究染色体的结构、遗传变异、基因定位、荧光原位杂交等奠定了基础。

[Objective] To explore the conditions of whole blood cell cultivation and chromosome preparation for Clarias leather.[Method] The blood was collected from the sterilized Clarias leather by disposable injection syringes with depo-heparin.The whole culture medium of RPMI1640（GIBCO） was used to incubate the blood cell in vitro.Through researching the cultivation temperature after getting blood from the Clarias leather,the content of colchicines and adding time,hypotonic treatment temperature and time and so on,a fairly mature experimental method was formed to cultivate the whole blood cell and chromosome preparation of Clarias leather.[Result] By adding 0.2 ml blood into 5 ml whole blood medium,cultivating under 26 ℃ for 72 h,and treating the blood cells with colchicines（final concentration of 0.1 μg/ml） 6 h before cultivation finished,and hypotonic treatment for 35min,better chromosome samples could be obtained.[Conclusion] The results laid foundation for further studying the structure,genetic variation,genetic mutation,and fluorescence in situ hybridization of chromosome.