人肾癌细胞原代培养方法比较及其生长特性观察

目的:分析比较三种方法分离人肾癌组织原代培养肾癌细胞及其传代能力，探讨建立理想的人肾癌细胞体外实验模型。方法:分别采用组织块贴壁法、机械分散法及差速贴壁分离法、混合酶消化法及差速贴壁分离法进行人肾癌细胞的体外培养，并从细胞形态学、表型、增殖生长曲线和细胞活力曲线等方面进行对比研究。结果:三种方法均可获得原代肾癌细胞，采用组织块贴壁法可获得较多数量的肾癌细胞，采用混合酶消化加分步贴壁后培养情况最好，数量较多，形态完整，第4代后肿瘤细胞状态稳定，增殖活跃，有明显的对数生长期，且与肾癌细胞系769-P细胞具有相似的表面抗原表达。结论:采用混合酶消化加分步贴壁法可获得较纯的肿瘤细胞，培养生长时间短，成功率高，适宜体外原代培养。

Comparison of primary culture methods of human kidney cancer cells and their growth characteristics observation

Objective:To compare the three methods to isolate human renal cell carcinoma primary cultured kidney cancer cells and passage ability,to establishment of ideal human kidney cancer cell model in vitro.Methods:Using the method of tissue adhesion,mechanical separation,mixed enzyme digestion and differential adherent separation to culture human renal cell carcinoma cells in vitro.Compare them from cell morphology,phenotype,proliferative growth curve and cell viability curve.Results:Theses three methods all could obtain primary renal cell carcinoma cells.Tissue adhesion method can get a larger number cells.But the mixed enzyme digestio could culture the best cells,which have complete cell morphology,at the fourth generation could be stable and proliferate actively,and have the similar surface antigen expression with 769-P cell lines.Conclusion:The mixed enzyme digestio could get more pure tumor cells,with shorter growth time,higher success rate,so it is suitable for primary culture in vitro.