title: [Isolation, culture and differentiation of skeletal muscle satellite cells of Luxi cattle embryo].
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: College of Animal Veterinary, Inner Mongolia Agricultural University, Hohhot 010018, China.
authors.forename: Yanchao
authors.initials: Y
authors.lastname: Bian
authors.forename: Xiaohong
authors.initials: X
authors.lastname: He
authors.forename: Ren
authors.initials: R
authors.lastname: Mu
authors.forename: Xiangchen
authors.initials: X
authors.lastname: Li
authors.forename: Yuehui
authors.initials: Y
authors.lastname: Ma
authors.forename: Yunzhang
authors.initials: Y
authors.lastname: Li
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002999:Clone Cells / Q000166:cytology; D015925:Cryopreservation; D004622:Embryo, Mammalian / Q000166:cytology*; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism