S180克隆细胞株的选择及其细胞特性研究

Clonal Cell Selection and Comparison Study of S180 Cell lines

目的 对北京市肿瘤研究所及本实验动物学部保存的S180进行了克隆培养，根据克隆细胞的腹水生长特性，从中选出8株克隆，对不同克隆S180细胞的特性进行研究。方法 免疫组化法测不同克隆细胞的蛋白质分布，流式细胞仪测定不同克隆细胞的DNA含量，扫描电镜观察克隆细胞表面结构，SDS-聚丙烯酰胺凝胶电泳测克隆细胞的电泳图谱。结果 其中3株克隆从可见腹水之日起，腹水即为血性；1株微显血性；4株在接种腹腔7日内腹水无血性。扫描电镜观察6株克隆细胞表面结构，各具特点，有所不同。流式细胞仪测定6株不同S180克隆细胞DNA含量，分别为8.5, 9.3，7.9，8.5，8.6，8.3pg。免疫组化染色显示不同克隆与11种抗体反应有所不同，一旦与某抗体反应阳性，则阳性细胞比率均在97%以上。SDS-聚丙烯酰胺凝胶电泳测定克隆细胞S1H10及S2D9的电泳图谱，S1H10克隆细胞有33条带，S2D9克隆细胞有30条带。 结论 8株克隆细胞各具特点。

Objective To isolate S180 stored in Beijing Cancer Institute and our department by the single cell cloning technique and to study property of those clonal cells. Methods SDS polyacrylamide gel electrophoresis, scanning electron microscopy and immunohistochemical method were used. Results Five sub-lines(S2H7,S1F11,S2D7,S2D9, S1B11)were isolated from S180 stored in Beijing Cancer Institute , three sub-lines (S1H10,S3F5, S1E11) were isolated from S180 stored in our department. Ascites of three sub-lines S1H10,S3F5 and S2H7 were bloody; S1F11 was tiny bloody; S1E11，S2D7，S2D9，S1B11 were not bloody porperty in 7 days of intraperitoneal inoculation.As for surficial structure under scanning electron microscope S3F5 had plentiful microvilli;S1B11 surface had a lot of prominences; S1F11 surficial microvilli were fewer than S3F5; S2D9 and S2H7 had different many tiny crests and tiny folds; S2D7 was more smooth.DNA contents were 8.5，9.3，7.9，8.5，8.6，8.3pg for six sublines S1B11，S2D7，S3F5，S2H7，S2D9，S1F11 respectively by Flow Cytometry. With immunohistochemistry the different clonal cells had different reactions with 11 kinds of antibodies (once reaction positive,positive rate was more than 97% ).S1H10 and S2D9 were detected by SDS-PAGE,with 33 and 30 protein bands respectively. Conclusion Eight sub-lines showed different properties.