Chapter title |
Primary Mouse Myoblast Purification using Magnetic Cell Separation
|
---|---|
Chapter number | 3 |
Book title |
Muscle Stem Cells
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6771-1_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6769-8, 978-1-4939-6771-1
|
Authors |
Marie Claude Sincennes, Yu Xin Wang, Michael A. Rudnicki, Sincennes, Marie Claude, Wang, Yu Xin, Rudnicki, Michael A. |
Editors |
Eusebio Perdiguero, DDW Cornelison |
Abstract |
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. |
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