MyoCult™ Expansion 10X Supplement (Mouse)
Serum-containing supplement for the expansion of mouse skeletal muscle progenitor cells (satellite cells)
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In-lab demonstration
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DMEM/F-12 with 15 mM HEPESDulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer
Overview
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).
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.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (5)
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
