Isolation,long-term culture,and ultrastructural characterization of adult cardiomyopathic cardiac muscle cells |
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Authors: | Asish C Nag Mei Cheng |
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Institution: | (1) Department of Biological Sciences, Oakland University, 48063 Rochester, Michigan |
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Abstract: | Summary A long-term cell culture system for adult cardiomyopathic hamster cardiac muscle cells has been established. The diseased
and control hearts were dissociated into single cell suspension with the modifications of our previous technique using collagenase
and hyaluronidase as applied to the dissociation of the adult rat heart. The postperfusion of the diseased heart with Krebs-Ringer
phosphate buffer and bovine serum albumin was very helpful in obtaining greater yield of viable diseased muscle cells; the
cells were cultured for 4 wk. Approximately 60% of the myocytes from the diseased heart and 85% of the myocytes from the normal
heart attached to the substrates and survived throughout the culture period. Approximately 60 to 70% of the cardiac myocytes
from the diseased and control hearts were bi- or multinucleated; 30% of the diseased and 80% of the normal myocytes showed
rhythmic contractility. Electron microscopy revealed the presence of two kinds of cardiac muscle cells in the diseased cell
culture on the basis of their myofibril content: one with scanty myofibrils and another with abundant myofibrils. Myocytes
with sparse myofibrils showed certain characteristic features that included autophagic vacuoles, amorphous matrix of fine
filamentous texture, scattered strips of myofibrils, and abnormal organization of the Z-line. Cardiac muscle cells with abundant
myofibrillar content contained unorganized myofibrils in certain sarcomeres. These studies demonstrate the feasibility of
maintaining diseased cardiac muscle cells from adult cardiomyopathic hamsters for at least 4 wk in monolayer culture.
This study was supported by a grant from the American Heart Association of Michigan, National Institutes of Health grant HL-25482,
and by an Oakland University Biomedical Research Support Grant. |
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Keywords: | cardiomyopathic hamster myocytes cell culture light microscopy electron microscopy |
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