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  • Isolation and Characterization of Adult Cardiac Fibroblasts and Myofibroblasts.

Isolation and Characterization of Adult Cardiac Fibroblasts and Myofibroblasts.

Journal of visualized experiments : JoVE (2020-04-01)
Meiling Melzer, David Beier, Pampee P Young, Sarika Saraswati
ABSTRACT

Cardiac fibrosis in response to injury is a physiological response to wound healing. Efforts have been made to study and target fibroblast subtypes that mitigate fibrosis. However, fibroblast research has been hindered due to the lack of universally acceptable fibroblast markers to identify quiescent as well as activated fibroblasts. Fibroblasts are a heterogenous cell population, making them difficult to isolate and characterize. The presented protocol describes three different methods to enrich fibroblasts and myofibroblasts from uninjured and injured mouse hearts. Using a standard and reliable protocol to isolate fibroblasts will enable the study of their roles in homeostasis as well as fibrosis modulation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-Actin, α-Smooth Muscle, clone 1A4, ascites fluid
Sigma-Aldrich
Anti-FSP1/S100A4 Antibody, from rabbit
Sigma-Aldrich
Monoclonal Anti-Vimentin antibody produced in mouse, clone LN-6, ascites fluid