Isolation, identification and biological characteristics of skeletal muscle satellite cells in pigeons

Objective  To investigate the method of isolation, culture and identification of white king pigeon(Columba livia) skeletal muscle satellite cells, and establish a complete culture system of pigeon skeletal muscle satellite cells.

Method  We selected 16-day pigeon embryos as experimental materials. Skeletal muscle satellite cells were isolated from the pectoral muscle by the tissue explants adherent method and collagenase digestion method, and the growth curve was drawn. MyHC expression was detected by immunofluorescence assay after differentiation of the satellite cells into myotubes. We detected mRNA expression of Desmin, Pax7, MyoG and MyoD1 genes before and after cell differentiation into myotubes using RT-qPCR.

Result  Skeletal muscle satellite cells were successfully isolated by the tissue explants adherent method and collagenase method, and cell growth appeared in S-shaped curve. After seven days of culture in high glucose DMEM containing 20% fetal bovine serum(FBS), a large number of visible myotubes were observed, and the myogenic differentiation marker MyHC was expressed in differentiated cells. RT-qPCR results showed that the relative expressions of Desmin and MyoG genes in differentiated myotube cells were 5.68 and 10.38 times of those in skeletal muscle satellite cells before differentiation, while the relative expressions of Pax7 and MyoD1 genes in skeletal muscle satellite cells before differentiation were 7.01 and 5.51 times of those in differentiated myotube cells, respectively.

Conclusion  The culture method of pigeon skeletal muscle satellite cells has been established, which provides a cell model for future studies of muscle development in pigeons.