Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
Nat Protoc. 2010 Apr;5(4):702-13. doi: 10.1038/nprot.2010.11. Epub 2010 Mar 25.
The successful isolation and cultivation of prostate stem cells will allow us to study their unique biological properties and their application in therapeutic approaches. Here we describe step-by-step procedures on the basis of previous work in our laboratory for the harvesting of primary prostate cells from adolescent male mice by a modified enzymatic procedure; the isolation of an enriched population of prostate stem cells through cell sorting; and the cultivation of prostate stem cells in vitro and characterization of these cells and their stem-like activity, including in vivo tubule regeneration. Normally, it will take approximately 8 h to harvest prostate cells, isolate the stem cell-enriched population and set up the in vitro sphere assay. It will take up to 8 weeks to analyze the unique properties of the stem cells, including their regenerative capacity in vivo.
成功分离和培养前列腺干细胞将使我们能够研究它们独特的生物学特性及其在治疗方法中的应用。在此，我们在实验室之前的工作基础上，描述了从青春期雄性小鼠中通过改良酶处理程序收获原代前列腺细胞、通过细胞分选分离富集的前列腺干细胞群体，以及在体外培养前列腺干细胞并对这些细胞及其类干细胞活性进行鉴定的分步程序，包括体内小管再生。通常，收获前列腺细胞、分离富含干细胞的群体和建立体外球体分析需要大约 8 小时，分析干细胞的独特特性，包括其体内再生能力，则需要 8 周时间。
