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PriCells: Isolation of Mice Muscle Cells
PriCells: Isolation of Mice Muscle Cells
The gastrocnemius, soleus, and plantaris were excised from three C57BL/6-Ly-5.1 6-week-old mice.
Tendons, all bone, and fat were carefully discarded, and the muscle tissue was thoroughly minced and then digested at 37°C with 0.2% collagenase for 45 min, followed by 0.1% trypsin for 45 min (PriCells Isolation Kit).
The tissue was triturated vigorously and passed through a 70-μm filter, and the cells were collected by centrifugation.
The cells were then plated in PriCells Culture System for 1 h at 37°C.
The nonadherent cells were then transferred to another plate, and the adherent cells (primarily fibroblasts) were discarded.
After 24 h, the floating cells and debris were washed off the plate, and fresh medium was applied to the attached cells.
After 5 days of culture, around 2 × 105 cells were collected from the plate after light trypsinization.