人子宫内膜腺癌细胞原代培养方法及细胞系鉴定

Primary culture of human endometrial adenocarcinoma cells and identification of endometrial adenocarcinoma cell lines

目的优化人子宫内膜癌细胞原代培养方法,建立体外培养细胞系。方法将辽宁医学院附属第一医院2009年10月至11月收治的3例子宫内膜癌患者手术切除的子宫内膜腺癌组织采用酶消化法制成单细胞悬液,用含血清DMEM/F-12培养基进行原代和传代培养,通过自然纯化、冻存及低血清培养法纯化腺上皮癌细胞。在倒置显微镜下观察细胞形态,行HE染色,免疫荧光染色对细胞进行鉴定,绘制生长曲线。结果 2例子宫内膜腺癌体外成功建系,传代数扩展至50～70代。细胞形态符合上皮癌细胞系特征,细胞角蛋白表达阳性,细胞生长曲线为"S"形。结论应用酶消化法原代培养子宫内膜腺癌细胞,并采用自然纯化、冻存及低血清培养法纯化腺上皮癌细胞简单、易行、高效。此2株细胞系可望作为子宫内膜腺癌研究的实验模型。

Objective To improve the method of primary culture of human endometrial adenocarcinoma cells and to establish cell lines of these ceils. Methods Tumor tissues from 3 patients with endometrial adenocarcinoma were made into single-cell suspension and cultured in DMEM/F-12 medium supplemented with fetal bovine serum. Purification of epithelial cells was attended by passage and cryopreservation. The morphology of the cells was observed under invert microscope and the cells were identified by immunofluorescence staining. The cell growth curve was plotted. Results We have successfully established 2 novel endometrial adenocarcinoma cell lines, which could be passaged for 50 or 70 generations with unchanged morphology and growth cycle. The morphology of cells indicated that the cell lines maintain the biologic characteristics of the malignant epithelial tumors. The cells were positively stained with cytokeratin. The cell growth curve showed sigmoid one. Conclusion The methods used in this study are convenient and of high performance. The established cell lines might be stable cell lines of endometrial adenocarcinoma.