Primary Mouse Myoblast Purification using Magnetic Cell Separation
Primary myoblasts can be isolated from mouse muscle cell extracts and cultured in vitro. Muscle cells are usually dissociated manually by mincing with razor blades or scissors in a collagenase/dispase solution. Primary myoblasts are then gradually enriched by pre-plating on collagen-coated plates, based on the observation that mouse fibroblasts attach quickly to collagen-coated plates, and are less adherent. Here, we describe an automated muscle dissociation protocol. We also propose an alternative to pre-plating using magnetic bead separation of primary myoblasts, which improve myoblast purity by minimizing fibroblast contamination.
Key wordsPrimary myoblasts Satellite cells Magnetic separation Fibroblast contamination Magnetic-activated cell sorting Muscle cells Pre-plating
Y.X.W. is supported by fellowships from QEII-GSST and the Canadian Institutes of Health Research. M.A.R. holds the Canada Research Chair in Molecular Genetics. These studies were carried out with support of grants to M.A.R. from the US National Institutes for Health [R01AR044031], the Canadian Institutes for Health Research [MOP-12080, MOP-81288], E-Rare-2: Canadian Institutes of Health Research/Muscular Dystrophy Canada [ERA-132935], the Muscular Dystrophy Association, and the Stem Cell Network.
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