The cells were cultured from atrium of the green fluorescent protein (GFP)-expressing male Sprague-Dawley rat (SD-Tg[CAG-EGFP]; Sankyo Lab, Tokyo, Japan) hearts. Under anesthesia, the heart was dissected and perfused with phosphate buffered saline (PBS; Wako, Tokyo, Japan) containing heparin sodium (Mochida Pharma, Tokyo, Japan) to wash out the blood. The atrium of the heart was next collected, cut into small pieces (less than 1 mm), and digested with 0.05% trypsin-ethylenediaminetetraacetic acid (EDTA; Sigma-Aldrich, Tokyo, Japan) for 9 min. These pieces were plated onto fibronectin-coated dishes (BD Biosciences, Tokyo, Japan) in Iscove’s modified Eagle’s medium (Life Technologies, Tokyo, Japan) supplemented with 20% fetal bovine serum (Thermo Scientific, Yokohama, Japan), 1% penicillin-streptomycin (Life Technologies, Tokyo, Japan). Two weeks later, the adherent outgrowth cells grew radially and were harvested to culture until second passage to expand the number of the cells.
Iscove s modified eagle s medium
Manufactured by Thermo Fisher Scientific
Sourced in Japan
Iscove's modified Eagle's medium is a cell culture medium formulation used to support the growth and maintenance of various cell types in laboratory settings. It provides the essential nutrients, vitamins, and other components required for cellular proliferation and survival.
Automatically generated - may contain errors
2 protocols using iscove s modified eagle s medium
1
Isolation and Culture of Atrial Cardiomyocytes from GFP-expressing Rats
2
Isolation and Culture of Atrial Cardiomyocytes from GFP-expressing Rats
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The cells were cultured from atrium of the green fluorescent protein (GFP)-expressing male Sprague-Dawley rat (SD-Tg[CAG-EGFP]; Sankyo Lab, Tokyo, Japan) hearts. Under anesthesia, the heart was dissected and perfused with phosphate buffered saline (PBS; Wako, Tokyo, Japan) containing heparin sodium (Mochida Pharma, Tokyo, Japan) to wash out the blood. The atrium of the heart was next collected, cut into small pieces (less than 1 mm), and digested with 0.05% trypsin-ethylenediaminetetraacetic acid (EDTA; Sigma-Aldrich, Tokyo, Japan) for 9 min. These pieces were plated onto fibronectin-coated dishes (BD Biosciences, Tokyo, Japan) in Iscove’s modified Eagle’s medium (Life Technologies, Tokyo, Japan) supplemented with 20% fetal bovine serum (Thermo Scientific, Yokohama, Japan), 1% penicillin-streptomycin (Life Technologies, Tokyo, Japan). Two weeks later, the adherent outgrowth cells grew radially and were harvested to culture until second passage to expand the number of the cells.
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