MyoCult™ Expansion 10X Supplement (Mouse)

Serum-containing supplement for the expansion of mouse skeletal muscle progenitor cells (satellite cells)
概要
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).
Advantages
• Supports robust expansion of mouse skeletal muscle progenitor cells.
• Culture-expanded skeletal muscle progenitor cells maintain differentiation potential.
• Rigorous raw material screening and quality control minimize lot-to-lot variability.
Subtype
Specialized Media
Cell Type
Myogenic Stem and Progenitor Cells
Species
Mouse
Application
Cell Culture, Expansion
Brand
MyoCult
Area of Interest
Stem Cell Biology
技术资料
数据及文献

Data

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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