Non-Enzymatic Isolation of Cancer-Associated Fibroblasts From Human Prostate Tumor Explants

Giulia Gangarossa

doi:10.21769/BioProtoc.5614

Non-Enzymatic Isolation of Cancer-Associated Fibroblasts From Human Prostate Tumor Explants

2026-03-05

0

PRE

AI

G

Giulia Gangarossa

C

Caterina Grillo

S

Sara Roccabianca

P

Pranzini, Erica

I

Iozzo, Marta

G

Giacomo Venditti

G

Greta Bertoli

I

Ippolito, Luigi *

G

Giannoni, Elisa

C

Comito, Giuseppina

C

Chiarugi, Paola

DOI：10.21769/BioProtoc.5614

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Abstract

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Prostate carcinoma (PCa) progression is strongly influenced by the surrounding tumor microenvironment, where cancerassociated fibroblasts (CAFs) represent the most abundant and functionally relevant stromal population. Despite their importance, the lack of stable cell lines representing CAF phenotypes limits the study of stromal-tumor interactions. To address this limitation, we provide an optimized protocol for isolating CAFs from fresh human PCa biopsies based on a mechanical procedure exploiting the specific CAF ability to migrate out from the tumor explants. This approach preserves tissue architecture and maintains CAF viability and phenotype. The resulting ex vivo CAF cultures provide a suitable model to investigate CAF biology within the tumor microenvironment.

Keywords:

Prostate cancer

Cancer-associated fibroblasts

Fibroblast activation

Primary cultures

Explant culture

Non-Enzymatic Isolation of Cancer-Associated Fibroblasts From Human Prostate Tumor Explants

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