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目的:建立小鼠宫颈癌肿瘤细胞系,观察其生物学特性。方法:采用肿瘤原代培养法对小鼠U14肿瘤进行体外传代培养,定期对培养细胞进行形态学观察,进行免疫组织化学鉴定、细胞周期检查、染色体检测、细胞生长测定,进行615、C57BL/c小鼠移植观察体内成瘤情况,并用pEGFP-N1质粒转染U14细胞。结果:细胞呈贴壁和悬浮混合生长,CK阳性特征。体外连续培养10个月,传代50代,细胞倍增时间为21.83小时,细胞周期测定G1期为34%,G2期为26.4%,S期为39.6%。培养U14细胞3-4天处于对数生长期,染色体为亚四倍体核型,众数为64～84条。615小鼠、C57BL/c小鼠移植成瘤率均为100%,无支原体污染。建立了GFP(+)的U14-GFP单克隆细胞株。结论:成功建立了小鼠U14子宫颈癌细胞系及GFP标记的U14-GFP细胞株,体内移植可建立常规及带荧光标记的肿瘤模型,可用于体内体外相结合的肿瘤研究。
Objective: To establish mouse cervical cancer cell lines and observe its biological characteristics. Methods: The primary tumor culture method was used to in vitro subculture the mouse U14 tumor. The cultured cells were observed morphologically on a regular basis. Immunohistochemistry, cell cycle test, chromosome test and cell growth assay were performed. The results of 615, C57BL / c The mice were transplanted to observe the in vivo tumorigenesis, and the U14 cells were transfected with pEGFP-N1 plasmid. Results: The cells were adherent and suspension mixed growth, CK positive features. The cells were cultured continuously for 10 months in vitro and passaged for 50 generations. The cell doubling time was 21.83 hours. The cell cycle was determined to be G1 at 34%, G2 at 26.4% and S at 39.6%. U14 cells cultured in 3-4 days in logarithmic growth phase, the chromosome sub-tetraploid karyotype, the mode is 64 to 84. 615 mice, C57BL / c mice transplanted into the tumor were 100%, no mycoplasma contamination. GFP (+) U14-GFP monoclonal cell line was established. Conclusion: The U14-GFP cell line and U14-GFP cell line have been successfully established in vivo. The conventional and fluorescent labeled tumor models can be established in vivo and can be used for oncology studies in vitro and in vivo.