简便绒毛细胞培养法制备染色体

目的探讨一种简便的绒毛细胞培养方法以及绒毛和胚芽染色体核型一致性分析。方法将33例标本(包括17例胚胎停育组和16例正常对照组)分别用胰蛋白酶法、胰蛋白酶加机械研磨法制备细胞悬液,培养后制备染色体,比较这两种消化方法在培养周期、核分裂相数等方面的差异;比较16例对照组标本在培养周期、分裂指数等方面的差异;将20例标本分别行绒毛、孕囊壁、胚芽染色体核型分析,比较三者染色体核型是否一致。结果 33例标本中有1例培养失败,培养成功率97%,16例胚胎停育组中检出异常核型7例。两种消化方法在绒毛细胞培养周期上具有显著相关性,但与核分裂相的数目无相关性。16例对照组中,与其他胎龄组相比,12周胎龄绒毛组织原代培养周期最长,有显著差异;二次传代周期和平均分裂指数均明显优于与原代。20例标本中有1例绒毛和胚芽染色体核型不一致。结论应用胰酶消化加机械研磨法制备绒毛细胞悬液,培养后制备染色体,方法简单高效;第二次传代培养有助于提高培养成功率并节约核型分析时间;对于同一标本,绒毛和胚芽染色体核型可能不完全一致。

A simple, convenient and practical method on in vitro primary cell culture of human villiform cells and chromosome preparation

Objective： To explore a simple, convenient and practical method on in vitro primary cell culture of human villiform cells and the chromosome karyotypic consistency of villiform ceils and embryonic bud. Methods ： 33 cases were divided into 17 cases of arrested intrauterine pregnancy in study group and 16 cases in normal control group. Each specimen of 33 cases was treated with trypsinase and trypsinase accompany with mechanical milling separately, villiform cells was harvested by the original culture to prepare chromosome. The culture cycle and the number of karyotypes were compared; 16 normal specimens of different pregnant weeks were compared on culture cycle and cell mitotic index; the villiform cells, embryonic bud and gestational sac of 20 specimens were cultured separately to prepare for chromosome. Results ： The time of villiform cell culture was obviously related to the kinds of digestive medthod, but not to the number of karyotypes. The villiform cells of 32 specimens （ 97% ） were cultured successfully and 7 abnormal karyotypes were tested in 16 cases of study groups. The culture cycle of the 12 weeks gestational age was the longest in the 16 cases of control groups compared with the others. The culture cycle and the Average cell mitotic index of second age were superior to those of the original culture. Among the 20 cases, only 1 case has different Chromosomal Karyotype between villiform and bud. Conclusion： The method of treating specimens with trypsinase and mechanical milling and preparing chromosome by the original cell culture is of high efficiency and favorable karyotypes ; the rate of success was enhanced by subculture ; Perhaps the chromosomal karyotype of villiform and bud in the same specimen are different.