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目的:探讨具有“永生化”特性的正常小鼠卵巢上皮细胞(MOSE)系建立的方法和可行性。方法:用胰酶消化法从温度敏感抗原(TsAg)转基因小鼠中分离MOSE,用4%血清培养液培养,通过细胞形态学特征的观察、细胞免疫组织化学染色检测CK-19和Vimentin来鉴别,体内成瘤实验观察有否恶性转化。结果:用胰酶消化法能迅速获得足够的MOSE,低血清(4%)培养可以有效去除可能污染的间质细胞,CK-19和Vimentin检测显示均符合上皮细胞的特性,细胞培养和体内成瘤实验均提示其符合具有“永生化”细胞的特性,没有自发发生恶性变,在体内没有成瘤能力。结论:用胰酶消化法,从转TsAg小鼠卵巢中获取的正常MOSE细胞具有“永生化”的特性,能够满足体外实验研究的需要,为卵巢癌的研究提供了可靠的细胞系。
Objective: To investigate the method and feasibility of establishing a mouse immortalized ovarian epithelial cell line (MOSE). Methods: MOSE was isolated from transgenic mice with temperature-sensitive antigen (TsAg) by trypsin digestion and cultured in 4% serum. Cell morphological characteristics were observed and CK-19 and Vimentin were detected by immunohistochemical staining , In vivo tumor experiment observed whether malignant transformation. RESULTS: Adequate MOSE was rapidly obtained by trypsin digestion, and low-serum (4%) culture was effective in removing possible contaminating interstitial cells. The results of CK-19 and Vimentin showed that all of them were in accordance with the characteristics of epithelial cells. Tumor experiments suggest that they are consistent with the characteristics of “immortalized” cells, no spontaneous malignant transformation, there is no tumor in vivo capacity. CONCLUSIONS: Normal MOSE cells obtained from the ovaries of TsAg-infected mice have the characteristics of “immortalization” by trypsin digestion, which can meet the needs of in vitro experimental studies and provide a reliable cell line for the study of ovarian cancer.