Purification and direct transformation of epithelial progenitor cells from primary human prostate

Justin M Drake, Andrew S. Goldstein, Daina L. Burnes, David S. Finley, Hong Zhang, Robert E. Reiter, Jiaoti Huang, Owen N. Witte

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

Epithelial cell transformation has been demonstrated in numerous animal models for the study of solid tumor biology. However, little evidence exists for human epithelial cell transformation without previous immortalization via genetic influences such as SV40 T-antigen, thus limiting our knowledge of the events that can transform naive human epithelium. Here we describe a system developed in our laboratory to directly transform freshly isolated primary human prostate epithelial cells without previous culture or immortalization. Prostate tissue is obtained from patients and benign tissue is separated from malignant tissue. Benign and malignant tissues are mechanically and enzymatically dissociated to single cells overnight, and immune cells and epithelial subsets are isolated on the basis of differential expression of surface antigens. Epithelial progenitor cells are transduced with lentiviruses expressing oncogenes and combined with inductive stroma for in vivo studies. At 8-16 weeks after transplantation into immune-deficient mice, the development of lesions, histologically classified as benign prostate, prostatic intraepithelial neoplasia and adenocarcinoma, can be evaluated.

Original languageEnglish (US)
Pages (from-to)656-667
Number of pages12
JournalNature Protocols
Volume6
Issue number5
DOIs
StatePublished - Apr 1 2011

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