MyoCult™ Expansion 10X Supplement (Mouse), when combined with DMEM/F-12 to prepare MyoCult™ Expansion Medium, is for the culture of mouse skeletal muscle progenitor cells (satellite cells). MyoCult™ Expansion Medium has been optimized for the expansion of mouse skeletal muscle satellite cells isolated by fluorescence-activated cell sorting (FACS), as well as for culturing single isolated myofibers grown in suspension. Satellite cells cultured using MyoCult™ Expansion Medium can be differentiated into multinucleated myotubes.
For preparation of MyoCult™ Expansion Medium, DMEM/F-12 with 15 mM HEPES (Catalog #36254) is required in addition to MyoCult™ Expansion 10X Supplement. For expansion of satellite cells, cultureware must be coated with a matrix such as Corning Matrigel® Basement Membrane Matrix (Corning Catalog #356234).
Subtype Specialized Media
Cell Type Myogenic Stem and Progenitor Cells
Species Mouse
Application Cell Culture, Expansion
Brand MyoCult
Area of Interest Stem Cell Biology
Data Figures
Figure 1. FACS-Isolated Pax7+ Mouse Satellite Cells Cultured in MyoCult™ Expansion Medium Expand More Efficiently Than in Homemade Medium
FACS-isolated CD45-/CD31-/Sca1-, alpha7-integrin+/Vcam1+ satellite cells were seeded at 2000 cells/well (24-well plate) and culture-expanded using complete MyoCult™ Expansion Medium (Mouse) or a commonly used homemade medium. Following 6 days of culture, (A) satellite cells were immunostained for nuclei (DAPI, blue) and Pax7 (red). Also, (B) total number and (C) percentage of Pax7+ satellite cells were quantified (n = 3). Error bars represent standard error of mean (SEM). Homemade medium was provided by the lab of Dr. Fabio Rossi, University of British Columbia.
Figure 2. FACS-Isolated Mouse Satellite Cells Are Expandable Over Multiple Passages in Myocult™ Expansion Medium
FACS-isolated CD45-/CD31-/Sca1-, alpha7-integrin+/Vcam1+ satellite cells were seeded at 2000 cells/well (24-well plate) and cultured using complete MyoCult™ Expansion Medium (Mouse) or a commonly used homemade medium. Following 12 days of expansion (2 passages), (A) satellite cells were imaged using phase contrast microscopy and (B) total numbers of MyoD+ satellite cells were quantified (n = 3). Error bars represent SEM. Homemade medium was provided by the lab of Dr. Fabio Rossi, University of British Columbia.
Figure 3. Mouse Satellite Cells Expanded in Myocult™ Expansion Medium Maintain Differentiation Capacity
FACS-isolated CD45-/CD31-/Sca1-, alpha7-integrin+/Vcam1+ satellite cells were culture-expanded using complete MyoCult™ Expansion Medium (Mouse) or a commonly used homemade medium for 2 passages (12 days following FACS isolation) and then treated with differentiation medium (high glucose DMEM with 2% Horse Serum). Following 4 days of differentiation, (A) myotubes cultured in MyoCult™ Expansion Medium were immunostained for nuclei (DAPI, blue) and Myosin Heavy Chain (MyHC, red), and (B) percentage of nuclei localized within MyHC+ myotubes (fusion index) was quantified (n = 3). Error bars represent SEM. Homemade medium was provided by the lab of Dr. Fabio Rossi, University of British Columbia.
Figure 4. Mouse Single Isolated Myofibers Can Be Cultured in MyoCult™ Expansion Medium Without Requiring Additional Supplements
Single isolated myofibers were cultured in suspension using complete MyoCult™ Expansion Medium (Mouse) or a homemade myofiber culture medium. After 4 days, (A) intact, viable myofibers were quantified (n = 3) and (B) immunostained for Pax7 (Red). Error bars represent SEM. Myofiber homemade medium was provided by the lab of Dr. Fabio Rossi, University of British Columbia. A specific myofiber medium (or additional supplements) is not needed for myofiber culture when using MyoCult™ Expansion Medium.
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