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J Vis Exp. 2015 Jul 20;(101):e52802. doi: 10.3791/52802.

Isolation and Characterization of Satellite Cells from Rat Head Branchiomeric Muscles.

Author information

1
Department of Orthodontics and Craniofacial Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center.
2
Department of Biological Structure, University of Washington School of Medicine.
3
Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center.
4
Department of Orthodontics and Craniofacial Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center; hans.vondenhoff@radboudumc.nl.

Abstract

Fibrosis and defective muscle regeneration can hamper the functional recovery of the soft palate muscles after cleft palate repair. This causes persistent problems in speech, swallowing, and sucking. In vitro culture systems that allow the study of satellite cells (myogenic stem cells) from head muscles are crucial to develop new therapies based on tissue engineering to promote muscle regeneration after surgery. These systems will offer new perspectives for the treatment of cleft palate patients. A protocol for the isolation, culture and differentiation of satellite cells from head muscles is presented. The isolation is based on enzymatic digestion and trituration to release the satellite cells. In addition, this protocol comprises an innovative method using extracellular matrix gel coatings of millimeter size, which requires only low numbers of satellite cells for differentiation assays.

PMID:
26274878
PMCID:
PMC4544364
DOI:
10.3791/52802
[Indexed for MEDLINE]
Free PMC Article

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