利用染色体G显带技术鉴定人永生细胞实验条件初探

Primary Exploration the Conditions of the Chromosome G-Banding Technique used in Examining the Human Immortal Lymphoblastoid Cells

目的：探索利用染色体G显带技术鉴定人永生淋巴细胞的最佳实验条件。方法：在常规染色体制片技术的方法和手段的基础上，分别观察不同PHA浓度和培养时间对永生淋巴细胞分裂增殖的影响；然后，选择不同浓度秋水仙素处理细胞，观察染色体分散程度和中期分裂相的形态。结果：在永生淋巴细胞培养48h，PHA终浓度为0．1mg／mL，秋水仙素终浓度为0．04μg/mL，且其作用时间为1．5～2h的条件下，所获得的细胞染色体标本分裂相较多、染色体较长、分散较好、利于带型分析。结论：利用染色体G显带技术鉴定人永生淋巴细胞提供了技术保证，降低了实验成本，并且缩短了实验时间。

Objective： To find best conditions in examining the human Immortal B lymphoblastoid ceils by the chromosome G - banding technology. Methods： Based on normal chromosome G- banding technology, we observed the factors which influence the cells＇ dispersion and the number of the metaphase fission phase, such as different concentration of PHA and colchicine and different culture time. Result： The human Immortal B lymphoblastoid cells were cultured for 48h with the final consistency of the PHA 0. lmg/mL, the ulthnate concentration of colchicine was 0.04μg/ mL and the reacting time＇ of it was 1.5 - 2h. In these conditions,we got more cells stopped at metaphase and chromosomes that were longer and dispersed enough to anylasis. Conclusion： The experiment gave us a best condition in examining the human Immortal B lymphoblastoid cells by the chromosome G - banding technology. Furthermore, the cost of the experiment was lowered and the time shorten.