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目的对肾透明细胞癌原代培养方法进行改进,以获得纯度及活力更高、遗传表型与来源更相近的肾透明细胞癌细胞系。方法将经过胶原酶消化法或机械法取得的肿瘤样本单细胞悬液,置入超低黏附培养板,用添加有丝分裂原等细胞因子的无血清培养基悬浮培养,离心滤除非肿瘤细胞和无成瘤能力肿瘤细胞,获得纯度高、异质性的肿瘤细胞囊球,囊球可以连续传代悬浮培养,也可转至常规培养条件培养,都可得到能连续传代的肿瘤细胞株。用流式细胞仪鉴定瘤球传代细胞,并与肾癌临床样本细胞进行表型比对。结果与单纯机械法或胶原蛋白酶消化法分离细胞及含血清培养基培养的方法相比,本法成功率更高,获得的肾透明细胞癌细胞保持与临床样本相似的异质性分化类型和表面抗原表达,并且无杂细胞污染。培养的细胞可经历稳定的连续传代并用于肿瘤相关研究。结论该方法简便易行,成功率高,培养的肾透明细胞癌细胞株可作为肾癌研究的良好实验模型。
OBJECTIVE To improve the method of primary culture of renal clear cell carcinoma in order to obtain a clear cell line with higher purity and vitality and more similar genetic phenotype and origin. Methods The single cell suspension of tumor samples obtained by collagenase digestion or mechanical method was placed in ultra-low adhesion culture plates, suspended in serum-free medium supplemented with mitogen and other cytokines, and centrifuged to filter non-tumor cells and non-tumor cells Tumor cells can obtain tumor cells with high purity and heterogeneity. The cyst balls and cyst balls can be continuously passaged and suspended in culture, and can also be transferred to conventional culture conditions to obtain tumor cell lines which can be continuously passaged. Cytometrocytoma cells were identified by flow cytometry and phenotypically aligned with clinical samples of renal cell carcinoma. Results Compared with the method of mechanically or collagenase digestion and culture of serum-containing medium, the success rate of this method was higher. The obtained clear cell carcinoma cells maintained similar heterogeneous differentiation types and surfaces as the clinical samples Antigen expression, and no contamination of the cells. The cultured cells can undergo stable serial passage and are used in tumor-related studies. Conclusion The method is simple and easy to operate with high success rate. The cultured renal clear cell carcinoma cell lines can be used as a good experimental model for renal cell carcinoma.