体外侵袭实验分离人肾癌细胞

In vitro separation of invasive subpopulation from human renal cell carcinoma

目的:探讨体外分离侵袭性和非侵袭性肾癌细胞的方法。方法:对肾细胞癌细胞株ACHN进行传代培养。在预实验确定铺胶浓度、消化回收时间的基础上,采用涂有Matrigel胶的Transwell对传代培养的肾癌细胞进行体外侵袭实验,分离回收侵袭性和非侵袭性肾癌细胞。结果:以浓度为1.17mg/ml、20μl/孔铺胶在Transwell,固定细胞悬液密度(5×106/ml),第48小时消化回收细胞较为理想。非侵袭性细胞呈散在生长,侵袭性细胞多呈聚集样生长。结论:Transwell可体外快速分离及回收、培养不同侵袭性的肾癌细胞群。用肾癌细胞株ACHN进行体外侵袭实验代表性好,且传代稳定性好。

Objective：To study the method for isolating the invasive and non-invasive cells from human renal cell carcinoma （RCC） cell line ACHN in vitro. Methods：The renal cell carcinoma cell line ACHN was serial subcultivated in vitro. Then in vitro invasion assay using the transwells coating matrigel were performed to separate and recover the invasive and non-invasive cells from the serial subculture. The concentration of matrigel, trypsinization and recovery time were subsequently optimized. Results： Matrigel （diluted into 1.17 mg/ml and 20 μl） was coated onto the filter of the Transwell. Cell suspension was prepared at a concentration of 5×10^5/ml and invasive cells were recovered after 48 h culture. When all above were prepared well, the recovery of invasive cells was performed ideally. The growth of non-invasive cells was sporadic, while that of the invasive cells was accumulative. Conclusion： In vitro the transwell is able to quickly separate, recover and culture the highly invasive RCC cells. At the same time, the cell line ACHN had a nice representation, and it can easily go down to the future generation.