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Aim : This study aimed to determine the effect of mechanical ventilation (MV) on the differentiation and proliferation of diaphragm satellite cells. Methods : Diaphragm satellite cells were isolated from C57 mice receiving 6 h of MV with optimized magnetic-activated cell sorting (MACS) approach. The cells were stained with BrdU or antibody for differentiation marker MYH3. The expression of MyoD and myogenin was detected by real-time PCR. Results : Diaphragm satellite cells were successfully isolated from mice by using MACS with a set of optimized parameters. About 1.5 × 10 5 cells could be harvested from a diaphragm. Upon MV, the proliferation rate of diaphragm satellite cells was decreased from 88.74% to 81.92%, while the differentiation rate was increased from 17.94% to 27.58%, compared to controls. Moreover, the expression of MyoD and myogenin were significantly upregulated upon MV. Conclusions : We established a practical method to purify diaphragm satellite cells, and demonstrated that MV regulated the differentiation and proliferation of diaphragm satellite cells.

Mechanical ventilation regulated the differentiation and proliferation of diaphragm satellite cells