[Isolation, identification, culture and bionomics of skeletal muscles satellite cells from green fluorescent protein transgenic mouse in vitro].
OBJECTIVE
To investigate the green fluorescent protein (GFP) expression and the bionomics of skeletal muscles satellite cells (SMSCs) in vitro in GFP transgenic mouse.


METHODS
The newborn transgenic mice were acquired to separate skeletal muscles satellite cells with enzyme digestion method. Cells were cultured and subcultured in vitro. Morphological observation, growth curve were investigated to evaluate the proliferation and differentiation characteristics of skeletal muscles satellite cells, fluorescence microscope was used to observe the GFP expression. The cells were identified by immunocytochemical stain. In the basis of identification of anti-sarcometric actin anti-body, the combination of anti-desmin antibody and DAPI (4, 6-diamidino-2-phenylindole) were used to detect the purification of skeletal muscles satellite cells.


RESULTS
Immunocytofluorescence suggested the good retain of GFP fluorescence in skeletal muscles satellite cells. The cells showed strong proliferative ability and they were positive with immunocytochemical stain of anti-sarcometric actin antibody and anti-desmin antibody. The combination of anti-desmin and DAPI stain can be used to determine the purification of SMSCs.


CONCLUSIONS
Skeletal muscles satellite cells cultured in vitro showed strong proliferation and differentiation ability. They are fit to construct the cell bank of tissure engineering and to be a useful tool to explore cells fate after transplantation since these cells retain the expression of GFP.
Dept. of Oral and Maxillofacial Surgery, Stomatological School of Shandong University, Jinan 250012, China.
Feng-He
FH
Zhang
Feng-Cai
FC
Wei
Ping
P
Huang
Shu-Yang
SY
Sun
Ke-Tao
KT
Wang
D000199:Actins; D000818:Animals; D001323:Autoantibodies; D002454:Cell Differentiation; D002478:Cells, Cultured; D003893:Desmin; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal; D032496:Satellite Cells, Skeletal Muscle
D000199:Actins; D001323:Autoantibodies; D003893:Desmin; C051932:anti-desmin antibody; D049452:Green Fluorescent Proteins