兴义维蚋蚋卵（胚胎）原代培养细胞核型分析

摘要

Objective:To culture primary cells of blackfly eggs( embryos)and analyze their karyo-type. Methods:The tissue block method was applied to treat the blackfly eggs( embryos)from Simu-lium( Wilhelmia)xingyiense that were collected in field. The blackfly eggs( embryos)were cultured in the modified Schneider′s and Sang M3 medium at pH 6 . 0~6 . 5 ,and incubated in a biochemical in-cubator at 28℃. The primary cells in the mitotic and proliferative phase were selected and treated with colchicine,and their metaphase chromosomes were prepared for karyotype analysis. Results:The pro-metaphase chromosomes are 3 pairs( 2 n=6 ),which were not aggregated at a maximum degree and showed themselves long rhabditiform,and the length of chromosome is 9. 43~13. 58 μm. Metaphase chromosome was 3 pairs( 2 n =6 ),and which were highly aggregated and showed themselves short rhabditiform,and the length of chromosome was 2. 43~4. 67 μm. Two metaphase chromosomes′arm ratio is less than 1. 67,belonging tocentric chromosome,and the 4 chromosomes′arm ratio is more than 1. 67,belonging to the submetacentric chromosome. Conclusions:The karyotype characteristics of primary cells cultured from the blackfly eggs( embryos)of Simulium( Wi. )xingyiense were prelimina-rily elucidated in this study.%目的：分析兴义维蚋蚋卵（胚胎）原代培养细胞核型。方法：应用组织块法处理野外采集兴义维蚋蚋卵（胚胎），以改良Shields and Sang M3为基础培养基，pH 6.0~6.5，置于28℃生化恒温培养箱中进行原代细胞培养，选取10个处于有丝分裂增殖时期的细胞，通过秋水仙素处理，制作中期染色体标本，进行核型分析。结果：显微镜下观察，前中期染色体为3对（2n=6），染色体凝集程度未达最大化呈现长杆状，染色体长度为9.43~13.58μm；中期染色体为3对（2n=6），高度凝集呈现短杆状，染色体长度为2.43~4.67μm，两条中期染色体臂比值<1.67，属中着丝粒染色体，4条染色体臂比值>1.67，属亚中着丝粒染色体。结论：初步阐析了兴义维蚋蚋卵（胚胎）原代培养细胞核型特征。