Purification and direct transformation of epithelial progenitor cells from primary human prostate.
Purification and direct transformation of epithelial progenitor cells from primary human prostate.
复制标题
纯化和直接转化上皮祖细胞从原代人前列腺中。
DOI：
10.1038/nprot.2011.317
复制
发表时间：
2011-05
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
--
作者：
上皮细胞转化已在许多用于实体瘤生物学研究的动物模型中得到证实。然而，几乎没有证据表明人类上皮细胞转化没有通过遗传影响（如SV40 T抗原）进行永生化，这限制了我们对可以转化幼稚人类上皮的事件的了解。在这里，我们描述了一个系统，在我们的实验室开发，直接转化新鲜分离的原代人前列腺上皮细胞，而无需事先培养或永生化。前列腺组织是从病人身上获得的，良性组织是从癌症中分离出来的。将良性和癌组织机械和酶促解离成单细胞过夜，并基于表面抗原的差异表达分离免疫细胞和上皮亚群。用表达癌基因的慢病毒转导上皮祖细胞，并与诱导基质组合用于体内研究。8-16在移植到免疫缺陷小鼠中5周后，可以评估组织学分类为良性前列腺、前列腺上皮内瘤（PIN）和腺癌的病变的发展。
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 prior immortalization via genetic influences such as SV40 T-antigen, limiting our knowledge of the events that can transform naïve human epithelium. Here we describe a system developed in our lab to directly transform freshly-isolated primary human prostate epithelial cells without prior culture or immortalization. Prostate tissue is obtained from patients, and benign tissue is separated from cancer. Benign and cancer tissue are mechanically and enzymatically dissociated to single cells overnight, and immune cells and epithelial subsets are isolated based on differential expression of surface antigens. Epithelial progenitor cells are transduced with lentiviruses expressing oncogenes and combined with inductive stroma for in vivo studies. 8-16 weeks after transplantion into immune-deficient mice, the development of lesions histologically classified as benign prostate, prostatic intraepithelial neoplasia (PIN) and adenocarcinoma can be evaluated.
登录
查看更多内容
DOI：
10.1126/science.1189992
发表时间：
2010-07-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Goldstein AS;Huang J;Guo C;Garraway IP;Witte ON
通讯作者：
Witte ON
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
50.3
作者：
Wang, SY;Gao, J;Wu, H
通讯作者：
Wu, H
影响因子：
11.2
作者：
Mulholland DJ;Xin L;Morim A;Lawson D;Witte O;Wu H
通讯作者：
Wu H
影响因子：
14.8
作者：
Tiscornia, Gustavo;Singer, Oded;Verma, Inder M.
通讯作者：
Verma, Inder M.