<i>Ex vivo</i> 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) <i>Ex vivo</i> Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples. Bio-protocol, 8 (21).

Access the full-text of this item by clicking on the Open Access link.

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 <i>ex vivo</i> 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 <i>ex vivo</i> prostate tissues, then assessed proliferation by immunohistochemistry of markers (H3S10<sup>phos</sup>). 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
Depositing User: Symplectic Admin
Date Deposited: 16 Nov 2018 11:06
Last Modified: 24 Mar 2021 13:10
DOI: 10.21769/bioprotoc.3075
Open Access URL: https://bio-protocol.org/e3075
URI: https://livrepository.liverpool.ac.uk/id/eprint/3028902