Primary Mouse Myoblast Purification using Magnetic Cell Separation.

Autor:	Sincennes MC; Sprott Center For Stem Cell Research, Ottawa Hospital Research Institute, Regenerative Medicine Program, 501 Smyth, Box 511, Ottawa, ON, Canada, K1H 8L6.; Faculty of Medicine, Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada., Wang YX; Sprott Center For Stem Cell Research, Ottawa Hospital Research Institute, Regenerative Medicine Program, 501 Smyth, Box 511, Ottawa, ON, Canada, K1H 8L6.; Faculty of Medicine, Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada., Rudnicki MA; Sprott Center For Stem Cell Research, Ottawa Hospital Research Institute, Regenerative Medicine Program, 501 Smyth, Box 511, Ottawa, ON, Canada, K1H 8L6. mrudnicki@ohri.ca.; Faculty of Medicine, Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada. mrudnicki@ohri.ca.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2017; Vol. 1556, pp. 41-50.
DOI:	10.1007/978-1-4939-6771-1_3
Abstrakt:	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.
Databáze:	MEDLINE
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