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https://hdl.handle.net/2440/124334
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Type: | Journal article |
Title: | Ex vivo culture and lentiviral transduction of Benign Prostatic Hyperplasia (BPH) samples |
Author: | McClurg, U.L. McCracken, S.R. Butler, L. Riabowol, K.T. Binda, O. |
Citation: | Bio-protocol, 2018; 8(21):e3075--e3075- |
Publisher: | Bio-protocol LLC |
Issue Date: | 2018 |
ISSN: | 2331-8325 2331-8325 |
Statement of Responsibility: | Urszula Lucja McClurg, Stuart R McCracken, Lisa Butler, Karl T Riabowol, Olivier Binda |
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 (H3S10phos). Herein, we describe how to generate lentiviral vectors and particules, successfully transduce human prostate tissues, induce exogenous gene expression, and assess cellular proliferation. |
Keywords: | Ex vivo tissue culture model; Proliferation; Oncogenic; Transformation; Prostate |
Rights: | Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC. |
DOI: | 10.21769/BioProtoc.3075 |
Published version: | http://dx.doi.org/10.21769/bioprotoc.3075 |
Appears in Collections: | Aurora harvest 4 Medicine publications |
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