Generating robust human valvular interstitial cell cultures: Protocol and considerations.
Cell culture
Valve disease
Valvular endothelial cell
Valvular interstitial cell
Journal
Journal of molecular and cellular cardiology
ISSN: 1095-8584
Titre abrégé: J Mol Cell Cardiol
Pays: England
ID NLM: 0262322
Informations de publication
Date de publication:
12 2022
12 2022
Historique:
received:
02
05
2022
revised:
19
10
2022
accepted:
23
10
2022
pubmed:
4
11
2022
medline:
30
11
2022
entrez:
3
11
2022
Statut:
ppublish
Résumé
Research in heart valve biology is a growing field that has yet to elucidate the fundamentals of valve disease. Human valvular interstitial cells (hVICs) are the best option for studying the cellular mechanisms behind valvular pathologies. However, there is a wide range of isolation procedures for these cells published in the literature. To what extent various isolation methods, patient pathologies, and seeding densities influence the behaviour of hVICs remains unclear. Here, we present an optimised method of hVIC isolation from diseased human valves donated at the time of surgery. We show that two rounds of 1000 U/mL collagenase digestion for not >2 h results in a phenotypically stable cell culture with a near complete absence of endothelial cell contamination. We also suggest that cells should be seeded at 10,000 cells/cm
Identifiants
pubmed: 36327771
pii: S0022-2828(22)00549-1
doi: 10.1016/j.yjmcc.2022.10.008
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
118-126Informations de copyright
Copyright © 2022 Elsevier Ltd. All rights reserved.