The aim of this study was to develop adequate in vitro conditions for the differentiation of bovine skeletal muscle cells. Therefore, satellite cells isolated from the left foreleg of a Holstein-Friesian fetus at 4.5 mo of gestation were seeded on 24-well plates coated with extracellular matrix gel. Cells were cultured for 5 d in growth medium containing 10% fetal bovine serum. After reaching confluence, several differentiation media were tested for inducing myotube formation. The highest fusion rate of approximately 30% was achieved with a serum-free medium containing 1 μM dexamethasone, 1 μg/ml linoleic acid, and 0.1 μM insulin after a differentiation phase of 72 h. Two different culture conditions (serum-free and serum-containing) appropriate for bovine skeletal muscle cell differentiation are described in detail which allow the investigation of bovine skeletal muscle cell proliferation and differentiation in general as well as in response to bioactive compounds.
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