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Myogenic satellite cells of Atlantic cod: development of an in vitro model to study muscle growth

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Kjetil Aune


Research Council's High North Conference; Tromsø, Norway, 2010-11-10–2010-11-11

Lazado, Carlo C.; Nagasawa, Kazue; Fernandes, Jorge

Muscle development is a complex phenomenon governed by diverse cellular mechanisms and molecular regulators. Myogenic satellite cells are self-renewing muscle stem cells and they proliferate at length to produce myoblasts that will later differentiate to form multinucleated myotubes. This study was aimed at isolating myogenic satellite cells from the fast myotomal muscle of juvenile Atlantic cod and characterizing their proliferation and differentiation in vitro. The myogenic satellite cells were isolated by a two-step enzyme digestion and were cultured in Dulbecco?s modified eagle?s media with NaHCO3, HEPES, horse serum and antibiotics. The cells were monitored daily and samples were taken at defined time-points for quantitative real-time PCR analysis of key gene markers for myogenic satellite cell growth and differentiation. Preliminary results showed that after 24h of culture spindle shaped morphology was exhibited by the cells. Intracellular fusion commenced at day 7 and at least 50% of the satellite cells virtually fused at day 20. Multinucleated myotubes started to be more evident at day 17 and distinct features were clearly observed at day 26. After a month of culture, an intricate web-like network was evidently observed in the cultures of myogenic satellite cells. The on-going study is on the expression of muscle-related genes which are key markers for myogenic satellite cells progression from quiescent state to proliferating and differentiating stage. Some of these genes are myod, pax7, desmin and gapdh. The results of the present study will elucidate how cod myogenic cells proliferate and differentiate in relation to expression of key myogenic genes. Furthermore, these myogenic cell cultures will be very useful in other downstream analyses such as the functional characterization of novel genes and testing of new substances.