[Isolation, culture and differentiation of skeletal muscle satellite cells of Luxi cattle embryo].

Objective: To isolate and culture skeletal muscle satellite cells of Luxi cattle embryo and study its biological characteristics.

Methods: The skeletal muscles were taken from the limbs of Luxi cattle embryo of 30-50 d old to isolate and culture the skeletal muscle satellite cells by differential adherence method and co-digestion of type I collagenase and trypsin. Satellite cell surface markers, desmin, MyoD, c-Met, Myf5 and pax7 were detected by immunocytochemistry and RT-PCR, and its biological characteristics were researched.

Results: The satellite cells were cultured to over passage 15. Cell viability was (97.90 ± 0.96)% after cryopreservation. The immunocytochemical staining showed that the specific surface antigen markers desmin and MyoD were positive, and RT-PCR also indicated the positive expressions of desmin, Myf5, c-Met and pax7. Colony formation was (56.39 ± 1.41)%. The karyotype analysis demonstrated that the isolated satellite cells were all derived from normal cattle embryos. Bovine satellite cells were successfully induced into osteoblasts and neuron-like cells by various inductors, and were positively expressed after alizarin red and toluidine blue staining. The osteoblast specific genes osteopontin and type I collagen and the nerve cell specific genes MAP-2 and nestin were positive by RT-PCR.

Conclusion: The skeletal muscle satellite cells of the Luxi cattle embryo are successfully isolated and cultured in vitro , They have the ability of being differentiated into osteoblasts and neuron-like cells.