Figure 4. PCA3-3STA detected primary prostate cancer cells from radical prostatectomy specimens ex vivo.

A. Scheme of the technique developed. Prostate biopsy cores are harvested from radical prostatectomy (surgery performed to treat PCa) or cystoprostatectomy (surgery performed to treat bladder cancer in which the benign prostate is removed as well) specimens immediately after surgical removal. The biopsy cores are then co-cultured with PCA3-3STA adenovirus during 24 h after which they are cultured for 48 h on a Gelfoam sponge. Three days after infection, the luciferase activity is measured using a CCD camera and is reported as p/s/cm² (Xenogen IVIS from Caliper Lifesciences). B. This procedure maintains PCa histological and specific biomarker expression such as prostate specific antigen (PSA), androgen receptor (AR), Ki67 or AMACR (positive in cancerous glands). C. Higher magnification of PCa histology and immunostainings as described in B). Note that the cytoplasmic AMACR staining is compatible with prostate cancer cells (e.g. see red arrows) while the nuclear stained cells correspond to p63 positive cells are found at biopsy periphery (see green arrow). Using this ex-vivo model, we show that PCA3-3STA-Luc adenovirus can detect primary PCa cells from radical prostatectomy specimens. Left panel: anti-firefly luciferase IHC, Middle panel: H&E staining; Right panel: anti-AMACR/p63 cocktail IHC.