成熟マウス心筋細胞の単離と模擬虚血モデルの作成

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Cultured cells are used for biochemical and physiological investigation of organ system, because they can supply relatively uniform experimental models. Cardiac myocytes have been also isolated and cultured for the use of heart studies. However, adult cardiac myocytes are sensitive to isolation condition and most studies utilized fetal or neonatal cardiac myocytes. Recently, several groups isolated adult cardiac myocytes from canine, rabbit and rat successfully. In this report, we tried to isolate myocytes from adult mice and to establish oxidative stress model of adult mouse myocytes. Hearts were excised and retro-perfused through aorta by collagenase solution following physiological salt solution containing uncoupler of excitation-contraction coupling, butanedione. Thereafter, hearts were minced in 1-2mm^3 cube and myocytes were isolated by mechanical agitation. Isolated cells were transferred onto culture dishes pre-coated with laminine. Cells were cultured with Dulbecco's modified Eagle medium containing 10% fetal bovine serum and bromodeoxyuridine. Isolated myocytes showed rod shape with striation, typical morphology of adult cardiac myocytes. They attached on laminine-coated dishes and survived over 5 days without proliferation of contaminated noncardiac myocytes. When cells were cultured more than a week, they changed their shape to spindle type and started spontaneous beating. To mimic oxidative stress to hearts, we added hydrogen peroxide to myocyte cultures. Myocytes released lactate dehydrogenase into culture medium dose and time dependently. These data suggest that we successfully isolated and cultured cardiac myocytes from adult mouse hearts. They showed similar characteristics to the heart in situ in oxidative stress model by hydrogen peroxide.
神戸女学院大学の論文
2003-07-20

成熟マウス心筋細胞の単離と模擬虚血モデルの作成

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