Leir Shih-Hsing, Browne James A, Eggener Scott E, Harris Ann
Human Molecular Genetics Program, Lurie Children's Research Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois; Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Section of Urology, University of Chicago Medical Center, Chicago, Illinois.
Fertil Steril. 2015 Mar;103(3):647-54.e1. doi: 10.1016/j.fertnstert.2014.11.022. Epub 2014 Dec 24.
To establish cultures of epithelial cells from all regions of the human epididymis to provide reagents for molecular approaches to functional studies of this epithelium.
Experimental laboratory study.
University research institute.
PATIENT(S): Epididymis from seven patients undergoing orchiectomy for suspected testicular cancer without epididymal involvement.
INTERVENTION(S): Human epididymis epithelial cells harvested from adult epididymis tissue.
MAIN OUTCOME MEASURE(S): Establishment of a robust culture protocol for adult human epididymal epithelial cells.
RESULT(S): Cultures of caput, corpus, and cauda epithelial cells were established from epididymis tissue of seven donors. Cells were passaged up to eight times and maintained differentiation markers. They were also cryopreserved and recovered successfully. Androgen receptor, clusterin, and cysteine-rich secretory protein 1 were expressed in cultured cells, as shown by means of immunofluorescence, Western blot, and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The distribution of other epididymis markers was also shown by means of qRT-PCR. Cultures developed transepithelial resistance (TER), which was androgen responsive in the caput but androgen insensitive in the corpus and cauda, where unstimulated TER values were much higher.
CONCLUSION(S): The results demonstrate a robust in vitro culture system for differentiated epithelial cell types in the caput, corpus, and cauda of the human epididymis. These cells will be a valuable resource for molecular analysis of epididymis epithelial function, which has a pivotal role in male fertility.
建立来自人类附睾所有区域的上皮细胞培养体系,为该上皮功能研究的分子方法提供试剂。
实验性实验室研究。
大学研究机构。
7例因疑似睾丸癌接受睾丸切除术且附睾未受累的患者的附睾。
从成人附睾组织中获取人类附睾上皮细胞。
建立成人人类附睾上皮细胞的稳健培养方案。
从7名供体的附睾组织中建立了头部、体部和尾部上皮细胞培养体系。细胞传代多达8次,并保持分化标记。它们也被成功冷冻保存和复苏。通过免疫荧光、蛋白质免疫印迹和定量逆转录聚合酶链反应(qRT-PCR)显示,培养的细胞中表达雄激素受体、簇集素和富含半胱氨酸的分泌蛋白1。通过qRT-PCR也显示了其他附睾标记物的分布。培养体系产生了跨上皮电阻(TER),其在头部对雄激素有反应,但在体部和尾部对雄激素不敏感,未刺激时体部和尾部的TER值要高得多。
结果表明建立了一种用于人类附睾头部、体部和尾部分化上皮细胞类型的稳健体外培养体系。这些细胞将是附睾上皮功能分子分析的宝贵资源,附睾上皮功能在男性生育中起关键作用。