Effect of skeletal muscle satellite cells C2C12 on the activity of chondrocytes ATDC5 in the co-cultured condition
Objective:To observe the effect of skeletal muscle satellite cells(MuSCs)C2C12 on the activity of chondrocytes ATDC5 in the case of co-cultured condition.Methods:①The C2C12 cells were cultured,and the third-generation cells were collected and divided into control group and dexamethasone(DEX)group.The C2C12 cells in the control group were cultured in the conventional Dulbecco's Modified Eagle's Medium(DMEM),while the ones in DEX group were intervened with DEX.After that,the effects of DEX on proliferation and pro-tein synthesis in C2C12 cells were determined by using the cell counting kit-8(CCK8)assay and bicinchoninic acid(BCA)assay,respec-tively,meanwhile,the wound scratch assay and Transwell chamber were employed for observing the effects of DEX on repairing and migra-tion ability of C2C12 cells.②The normal C2C12 cells(co-cultured group)and DEX-pretreated C2C12 cells(pre-treatment group)were co-cultured with ATDC5 chondrocytes,respectively,in the Transwell chambers;In the Transwell chamber of control group,the upper chamber was the cells-free conventional culture medium,and the lower was inoculated with ATDC5 cells.After that,the proliferation rate and intra-cellular reactive oxygen species(ROS)level of ATDC5 cells were detected by using CCK8 assay and 2',7'-dichlorofluorescin diacetate(DCFH-DA)fluorescent probe,respectively.Results:①The proliferation rate and protein level of C2C12 cells were lower in DEX group compared to control group(78.402±5.401 vs 100.000±3.096％,t=8.498,P=0.000;5 080.367±296.657 vs 5 775.577± 150.476 μg/mL,t=3.620,P=0.022).②The wound repairing rate of C2C12 cells was lower in DEX group in contrast to control group(53.173±1.800 vs 79.979±10.176％,t=4.493,P=0.011).③On hour 24 and 48,the number of migrated C2C12 cells was less in DEX group compared with that of control group(on hour 24:24.200±5.630 vs 57.000±2.449 cells,t=11.945,P=0.000;on hour 48:57.600±8.820 vs 91.000±4.743 cells,t=7.457,P=0.000).④There was statistical difference in proliferation rate of ATDC5 cells among the 3 groups(100.000±1.663,116.894±7.917,89.130±2.980％,F=23.700,P=0.001).The proliferation rate of ATDC5 cells was higher in control group and co-cultured group compared to pre-treatment group(P=0.037,P=0.006),and was highest in co-cultured group(P=0.001).⑤There was statistical difference in intracellular ROS level of ATDC5 cells among the 3 groups(20.148± 5.636,13.959±4.110,40.691±3.146,F=30.096,P=0.001).The intracellular ROS level of ATDC5 cells was higher in pre-treatment group compared to control group and co-cultured group(P=0.001,P=0.000),while,the difference between control group and co-cultured group was not statistically significant(P=0.137).Conclusion:DEX can inhibit the proliferation of C2C12 cells,and the growth state of MuSCs under the condition of muscle atrophy in vitro can be simulated by intervening C2C12 cells with DEX.In the case of normal condi-tions,the metabolic products of C2C12 cells can promote the proliferation of chondrocytes ATDC5,and the reduction in viability of C2C12 cells can inhibit the proliferation of chondrocytes ATDC5,as well as increase the oxidative damage to chondrocytes ATDC5.
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维普
