Ex vivo Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples

McClurg, Urszula Lucja ORCID: 0000-0003-2631-4174, McCracken, Stuart R, Butler, Lisa, Riabowol, Karl T and Binda, Olivier (2018) Ex vivo Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples BIO-PROTOCOL, 8 (21). e3075-. ISSN 2331-8325, 2331-8325

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Abstract

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 (H3S10^phos). Herein, we describe how to generate lentiviral vectors and particules, successfully transduce human prostate tissues, induce exogenous gene expression, and assess cellular proliferation.

Item Type:	Article
Uncontrolled Keywords:	Ex vivo tissue culture model, Proliferation, Oncogenic, Transformation, Prostate
Depositing User:	Symplectic Admin
Date Deposited:	16 Nov 2018 11:06
Last Modified:	22 May 2026 21:43
DOI:	10.21769/BioProtoc.3075
Open Access URL:	https://bio-protocol.org/e3075
Related Websites:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3028902
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