摘要
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/6j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21.83h,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/6j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 a murine uterine cervical cancer cell line and to define its biological characteristics. Methods： Transplanted tumor tissues were used for primary culture of U14 cervical carcinoma cells in vitro. After 20 passages, we detected CK expression with immunohistochemistry, observed cell cycle with flow eytometry, analyzed the chromosomes, determined the cell growth curve, and checked tumorigenicity and metastatic potential. We also transfected U14 cells with the plasmid pEGFP-N1. Results： The newly established murine cell line was continuously passaged 50 times in 10 months. The cells showed positive CK expression. The karyotype was hypotetraploid, and the modal number was 64-68. The cell doubling time was 21.83 hours, and the cell line shows exponential growth on the third and fourth days. Cell cycle analysis showed that 34% of the cells were in G1 phase, 26.4% of the cells were in G2 phase, and 39.6% of the cells were in S phase. After implantation, tumorigenicity was 100%. No mycoplasma was detected. We also established a monoclonal continuous cell strain named U14-GFP with 100% positive expression of GFP. Conclusion： We successfully established a new murine cervical carcinoma cell line named U14 and a related cell strain U14-GFP that expresses GFP. Both strains can be used for research in vivo or in vitro.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2008年第7期391-394,共4页
Chinese Journal of Clinical Oncology
基金
国家科技基础条件平台建设基金资助(编号:20051KA21502)
关键词
小鼠宫颈癌
细胞系
生物学特性
Mouse uterine cervical cancer
Cell line
Biological characteristic
