Effect of Oxygen and Glucose Concentration on In-vitro Development of Skeletal Muscle Satellite Cells

Objective:The effects of oxygen and glucose on the development of skeletal muscle satellite cells were evaluated in vitro, to analyze the reasons for proliferation and injury and repair of SCs.Methods:The primary passage SCs were derived from gastrocnemius muscle and soleus muscle of the Sprague-Dawley rats (4weeks of age, male) .The SCs were digested with 0.1% Collagenase II and 0.25% Trypsin, purified with means of differential attachment technique and identified withα-sarcometricactin.The third passage SCs were treated with different doses both of oxygen and glucose:21%O2&5.5mM Glu, 21%O2&25mM Glu, 1%O2&5.5 mM Glu, 1%O2&25mM Glu.The cell proliferation rate was measured by CCK-8cell proliferation assay after 3hand the levels of lactate dehydrogenase (LDH) released to the extracellular bathing media had been used for assessing SCs injury.Results:The experimental group of 21% O2&25mM Glu was with a higher proliferation rate than the experimental group of 21% O2&5.5mM Glu (P<0.01) .The experimental group of 1%O2&25mM Glu was with a higher proliferation rate than the experimental group of 1%O2&5.5mM Glu (P<0.01) .The experimental group of 1%O2&5.5mM Glu was with a higher proliferation rate than the experimental group of 21%O2&5.5mM Glu (P<0.01) . (2) The LDH level of experimental group of 1%O2&5.5mM Glu was higher than experimental group of 21%O2&5.5mM Glu (P<0.01) .The LDH level of experimental group of 1%O2&25mM Glu was higher than experimental group of 21%O2&25mM Glu (P<0.01) .Conclusion:Under the same oxygen condition, SCs' proliferation rate in DMEM of high-glucose was higher than that in DMEM of low-glucose.Under the same glucose concentration, SCs' proliferation rate in hypoxia was higher than that in normal oxygen.Under the same glucose concentration, the level of LDH released was higher in hypoxia than that in normal.