Co-cultures of Macrophages with Muscle Stem Cells with Fibroadipogenic Precursor Cells from Regenerating Skeletal Muscle

Adult muscle stem cells rebuild myofibers after damage. Although they are highly powerful to implement the adult myogenic program, they need environmental cues provided by surrounding cells for efficient and complete regeneration. Muscle stem cell environment includes fibroadipogenic precursors, vascular cells, and macrophages. A way to decipher the complexity of the interactions muscle stem cells establish with their neighborhood is to co-culture cells freshly isolated from the muscle and assess the impact of one cell type on the behavior/fate of the other cell type. Here, we present a protocol allowing the isolation of primary muscle stem cells, macrophages, and fibroadipogenic precursors by Fluorescence Activated Cell Sorting (FACS) or Magnetic Cell Separation (MACS), together with co-culture methods using a specific setup for a short time window to keep as much as possible the in vivo properties of the isolated cells.