TY - JOUR
T1 - Culture and Lentiviral Transduction of Benign Prostatic Hyperplasia (BPH) Samples
AU - McClurg, Urszula Lucja
AU - McCracken, Stuart R
AU - Butler, Lisa
AU - Riabowol, Karl T
AU - Binda, Olivier
PY - 2018/11/5
Y1 - 2018/11/5
N2 - 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.
AB - 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.
KW - Journal Article
M3 - Article
C2 - 30515449
SN - 2331-8325
VL - 8
JO - JMIR research protocols
JF - JMIR research protocols
IS - 21
ER -