摘要
目的 探讨大鼠良性增生前列腺腺上皮细胞系的建立方法. 方法 2011年12月至2012年12月选取13周龄雄性自发性高血压大鼠20只,屏障环境下常规饲养至29周龄.取大鼠前列腺组织行苏木精-伊红染色鉴定前列腺上皮增生情况.取前列腺腹侧叶组织,剪碎后用Ⅰ型胶原蛋白酶消化,收集细胞,分别使用WAJC-404、PrEGM培养液进行体外培养14 d并传代.免疫细胞化学方法（CK8/18）鉴定原代培养的增生前列腺腺上皮细胞的特异性表达.绘制细胞生长曲线,比较两种培养基培养前列腺腺上皮细胞的生长情况. 结果 采用WAJC-404、PrEGM培养基体外原代培养自发性高血压大鼠前列腺腺上皮细胞均达14 d,并成功纯化和传代.免疫细胞化学实验证实原代培养的增生前列腺腺上皮细胞特异性表达细胞角蛋白CK8和CK18.细胞生长曲线显示：PrEGM培养基培养的腺上皮细胞与WAJC-404培养基相比,细胞形态更好,细胞活力更为旺盛,进入对数生长期的时间更短（4d与7d）,细胞生长曲线平均峰值更高（15.3× 104/ml与12.8× 104/nl）. 结论 大鼠良性增生前列腺腺上皮细胞系可体外构建,PrEGM培养基比WAJC-404培养基更适合该细胞系的建立.
Objective To set up the methods of establishing rat primary benign prostatic hyperplasic glandular epithelial cell line.Methods Male spontaneously hypertensive rats were raised to 29 weeks,and then evaluated the situation of BPH with HE staining.The prostate tissue from ventral prostate lobe was aseptically removed,dissected,minced,and then dissociated in collagenase type Ⅰ.Isolated cells were collected,seeded in WAJC-404 and PrEGM medium separately,then cultured and passaged.Specificity of primitive cultured prostatic epithelial cells was identified by cell immunochemistry with CK8/18,and the cell growth curves were drawn.Then the situation of growth of the two prostatic hyperplasic glandular epithelial cell lines were analysed and compared.Results The prostatic hyperplasic glandular epithelial cell lines of the spontaneously hypertensive rats in WAJC-404 and PrEGM medium were successfully primarily cultured,purified and passaged in vitro.Cell immunochemistry proved that the cell lines specifically express cytokeratin 8/18.Cell growth curve showed that prostatic epithelial cells in PrEGM,compared with prostatic epithelial cells in WAJC-404,possessed better cell morphology,more exuberant cell vitality,faster growth rate to enter the logarithmic growth period（4 d vs.7 d）and higher peak of cell growth curve（15.3× 104/ml vs.12.8×104/ml）.Conclusions Rat primary benign prostatic hyperplasic glandular epithelial cell line can be established conventionally in vitro.PrEGM medium is more suitable for primary culture of the rat benign prostatic hyperplasic glandular epithelial cell line than WAJC-404 medium.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2014年第5期383-387,共5页
Chinese Journal of Urology
基金
国家自然科学基金（81070601）
关键词
前列腺腺上皮细胞
良性前列腺增生
细胞系
原代培养
Prostatic glandular epithelial cell
Benign prostatic hyperplasia
Cell line
Primary culture
