Isolation and culture of the terminally differentiated adult mammalian ventricular cardiac muscle cell |
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Authors: | William C Claycomb Nicholas Lanson Jr |
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Institution: | (1) Department of Biochemistry, Louisiana State University School of Medicine, 70112 New Orleans, Louisiana |
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Abstract: | Summary We have carried out systematic studies to optimize and standardize methodology to isolate and culture the adult rat ventricular
cardiac muscle cell. Four hearts were perfused simultaneously with a calcium-free medium containing collagenase. The ventricular
tissue was then minced and further digested to liberate individual cells. Approximately 16 million rod-shaped muscle cells
were obtained. The plating efficiency has been greatly improved by culturing the cells in a conditioned medium prepared from
a rabbit corneal cell line. This medium also contained added fetal bovine serum, essential and nonessential amino acids, vitamins,
insulin, transferrin, and 25 trace minerals. The culture flasks were precoated with rat-tail collagen. Fibroblast contamination
was virtually eliminated by including cytosine arabinoside in the medium during the first 7 d of culture. After this time
the cells could be cultured in the absence of serum in a chemically defined medium composed of MEM, vitamins, nonessential
amino acids, and trace minerals. They continued to contract spontaneously and do well in this medium for at least 3 d thereafter.
This improved methodology resulted in a reproducible culture system with improved plating efficiency. It provided a new and
unique system to study the structure and function of the adult mammalian ventricular cardiac muscle cell.
This investigation was supported by Grant HL 25873 from the National Institutes of Health, Bethesda, MD. |
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Keywords: | cardiac muscle adult rat cell culture heart methodology ventricular |
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