离体
前列腺
转导(生物物理学)
生物
细胞培养
癌症研究
基质
免疫组织化学
增生
细胞生长
体内
病毒载体
细胞生物学
病理
分子生物学
基因
免疫学
医学
重组DNA
癌症
遗传学
内分泌学
生物化学
作者
Urszula L. McClurg,Stuart McCracken,Lisa M. Butler,Karl Riabowol,Olivier Binda
出处
期刊:Bio-protocol
[Bio-Protocol]
日期:2018-11-05
卷期号:8 (21)
被引量:3
标识
DOI:10.21769/bioprotoc.3075
摘要
To assess oncogenic potential, classical transformation assays are based on cell line models. However, cell line based models do not reflect the complexity of human tissues. We thus developed an inducible expression system for gene expression in ex vivo human tissues, which maintain native tissue architecture, such as epithelia and stroma. To validate the system, we transduced and expressed known tumor suppressors (p53, p33ING1b), oncoproteins (RasV12, p47ING3), or controls (empty vector, YFP) in ex vivo prostate tissues, then assessed proliferation by immunohistochemistry of markers (H3S10phos). Herein, we describe how to generate lentiviral vectors and particules, successfully transduce human prostate tissues, induce exogenous gene expression, and assess cellular proliferation.
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