The University of Southampton
University of Southampton Institutional Repository

Isolation and characterization of biliary epithelial and stromal cells from resected human cholangiocarcinoma: a novel in vitro model to study tumor-stroma interactions

Isolation and characterization of biliary epithelial and stromal cells from resected human cholangiocarcinoma: a novel in vitro model to study tumor-stroma interactions
Isolation and characterization of biliary epithelial and stromal cells from resected human cholangiocarcinoma: a novel in vitro model to study tumor-stroma interactions
Cholangiocarcinoma (CCA) is a devastating malignancy arising from the bile ducts. Cancer-associated fibroblasts (CAFs) are key players in CCA invasiveness and in the generation of a desmoplastic reaction. The aim of the present study was to develop a novel model by which to study tumor-stroma interactions using primary cultures of human biliary epithelial cells (hBECs) and stromal cells (SCs) in CCA. hBECs and SCs, isolated from surgical resections (n=10), were semi-purified by centrifugation on a Percoll gradient; hBECs were further immunopurified. hBECs and SCs were characterized using epithelial [cytokeratin 7 (CK7) and CK19] and mesenchymal [vimentin (VMN), ?-smooth muscle actin (?-SMA), CD68] cell markers. The purity of cultured cells was assessed by fluorescent immunocytochemistry. hBECs were HEA125/CK7/CK19-positive and VMN/?-SMA-negative. SCs were VMN/?-SMA-positive and CK7/CK19-negative. CCA 2-D culture models have been described but they use long-standing CCA cell lines of various biliary tumor cell origins with stromal cells derived from non-cholangiocarcinoma tissues. Recently, a novel 3-D organotypic co-culture model of rat cholangiocarcinoma was described. In the present study, we obtained pure and stable primary cultures of hBECs and SCs from CCA surgical specimens. These cell cultures may provide a useful tool by which to study CCA tumor-stroma interactions
1021-335X
1143-1148
Massani, Marco
32fad209-0241-4168-8b07-06d17808a2b8
Stecca, Tommaso
196ca669-5540-4216-883d-ed9b32ee5ac8
Fabris, Luca
7ab16fef-75b0-4131-a451-86fc97903b92
Caratozzolo, Ezio
54f55bd0-af51-4b51-a482-a9ac6ec84845
Ruffolo, Cesare
e889b2df-cfaf-4ad5-b1ab-21d7c33f47eb
Furlanetto, Alberto
d909992a-86f2-4645-b3bc-18037494cd81
Morton, Stuart
7545cf7d-15a6-43cc-9949-41ece1749fef
Cadamuro, Massimiliano
eee3861c-58b8-43a4-99d2-24f488b45eae
Strazzabosco, Mario
18e0c674-2971-4029-8c9f-7770f14ea564
Bassi, Nicolò
0b829168-b99e-4258-8cf6-9f7136cc9d58
Massani, Marco
32fad209-0241-4168-8b07-06d17808a2b8
Stecca, Tommaso
196ca669-5540-4216-883d-ed9b32ee5ac8
Fabris, Luca
7ab16fef-75b0-4131-a451-86fc97903b92
Caratozzolo, Ezio
54f55bd0-af51-4b51-a482-a9ac6ec84845
Ruffolo, Cesare
e889b2df-cfaf-4ad5-b1ab-21d7c33f47eb
Furlanetto, Alberto
d909992a-86f2-4645-b3bc-18037494cd81
Morton, Stuart
7545cf7d-15a6-43cc-9949-41ece1749fef
Cadamuro, Massimiliano
eee3861c-58b8-43a4-99d2-24f488b45eae
Strazzabosco, Mario
18e0c674-2971-4029-8c9f-7770f14ea564
Bassi, Nicolò
0b829168-b99e-4258-8cf6-9f7136cc9d58

Massani, Marco, Stecca, Tommaso, Fabris, Luca, Caratozzolo, Ezio, Ruffolo, Cesare, Furlanetto, Alberto, Morton, Stuart, Cadamuro, Massimiliano, Strazzabosco, Mario and Bassi, Nicolò (2013) Isolation and characterization of biliary epithelial and stromal cells from resected human cholangiocarcinoma: a novel in vitro model to study tumor-stroma interactions. Oncology Reports, 30 (3), 1143-1148. (doi:10.3892/or.2013.2568). (PMID:23807641)

Record type: Article

Abstract

Cholangiocarcinoma (CCA) is a devastating malignancy arising from the bile ducts. Cancer-associated fibroblasts (CAFs) are key players in CCA invasiveness and in the generation of a desmoplastic reaction. The aim of the present study was to develop a novel model by which to study tumor-stroma interactions using primary cultures of human biliary epithelial cells (hBECs) and stromal cells (SCs) in CCA. hBECs and SCs, isolated from surgical resections (n=10), were semi-purified by centrifugation on a Percoll gradient; hBECs were further immunopurified. hBECs and SCs were characterized using epithelial [cytokeratin 7 (CK7) and CK19] and mesenchymal [vimentin (VMN), ?-smooth muscle actin (?-SMA), CD68] cell markers. The purity of cultured cells was assessed by fluorescent immunocytochemistry. hBECs were HEA125/CK7/CK19-positive and VMN/?-SMA-negative. SCs were VMN/?-SMA-positive and CK7/CK19-negative. CCA 2-D culture models have been described but they use long-standing CCA cell lines of various biliary tumor cell origins with stromal cells derived from non-cholangiocarcinoma tissues. Recently, a novel 3-D organotypic co-culture model of rat cholangiocarcinoma was described. In the present study, we obtained pure and stable primary cultures of hBECs and SCs from CCA surgical specimens. These cell cultures may provide a useful tool by which to study CCA tumor-stroma interactions

Full text not available from this repository.

More information

Published date: September 2013
Organisations: Medical Education

Identifiers

Local EPrints ID: 382372
URI: http://eprints.soton.ac.uk/id/eprint/382372
ISSN: 1021-335X
PURE UUID: 733ea166-98ed-4246-8dfe-cdf551a52f1f
ORCID for Stuart Morton: ORCID iD orcid.org/0000-0002-5921-4869

Catalogue record

Date deposited: 30 Oct 2015 09:38
Last modified: 20 Jul 2019 00:33

Export record

Altmetrics

Contributors

Author: Marco Massani
Author: Tommaso Stecca
Author: Luca Fabris
Author: Ezio Caratozzolo
Author: Cesare Ruffolo
Author: Alberto Furlanetto
Author: Stuart Morton ORCID iD
Author: Massimiliano Cadamuro
Author: Mario Strazzabosco
Author: Nicolò Bassi

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×