Investigating the Vascular Niche: Three-Dimensional Co-culture of Human Skeletal Muscle Stem Cells and Endothelial Cells
Angiogenesis, the growth of new blood vessels, is crucial for efficient skeletal muscle regeneration. Myogenesis and angiogenesis take place concomitantly during muscle regeneration. Myogenic precursor cells (MPCs) are in close proximity to vessels and interact with neighboring endothelial cells (ECs) to expand and differentiate. To demonstrate functional interplay between the two cell types, we established a robust and predictive ex vivo assay to evaluate activity of MPCs on angiogenesis and vice-et-versa, of ECs on myogenesis. Here, we describe an optimized three-dimensional co-culture protocol for the assessment of biological interactions between MPCs and ECs during skeletal muscle regeneration.
Keywords3D tri/co-culture Angiogenesis Endothelial cells Fibrin gel Muscle stem cells Myogenesis Niche
We thank Stéphane Germain and Laurent Muller (CIRB, Collège de France, Paris) for the transfer of the 3D angiogenesis technique in our lab. B. Chazaud group research using this technique has been recently supported by INSERM, CNRS, Université Claude Bernard Lyon I, and grants from the Framework Programme FP7 Endostem (grant agreement 241440), and Association Française contre les Myopathies (grants 18003 and 111199). C.L. was supported by Dim Stem Pole from Region Ile-de-France and Association Française contre les Myopathies.
- 11.Nakatsu MN, Davis J, Hughes CCW (2007) Optimized fibrin gel bead assay for the study of angiogenesis. J Vis Exp. https://doi.org/10.3791/186