• Ready-to-use, differentiated myotubes typically within 48 hours of thawing
  • No specialty medium required—just plate in DMEM with 2% horse serum
  • Validated for differentiation capacity—myogenic index greater than 50% after 2 days

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Skeletal muscle plays an essential role in maintaining physiological and metabolic health in humans. GIBCO® Human Skeletal Myoblasts (HSkM) offer an easy-to-use, cost effective cell system for studying conditions associated with loss of muscle mass and glucose uptake. Cells are thawed, plated in DMEM plus 2% horse serum, and are ready to use typically within 48 hours (Figure 1). These primary cells have been shown to respond to physiological concentrations of ligands including TGF-β1 and Myostatin and thus are more relevant than commonly used rodent models (Figure 2).

Figure 1. GIBCO® Human Skeletal Myoblasts (HSkM) are easy to use, and do not require counting— just dilute in the specified volume of medium and plate.

Cells are thawed and plated in DMEM supplemented with 2% horse serum. Cells differentiate typically within 48 hours and form myotubes that are ready for various applications. The system is compatible with multi-well formats from 6-well to 384-well plates. The top right panels show differentiated cells with bound anti-troponin mAb (clone CT3), detected with an anti-mouse Alexa Fluor® 546 secondary antibody (Cat. No. A11030) and Hoechst 33342 nuclear counterstain (Cat. No. H3570).

Figure 2. Physiologically relevant cellular model.

GIBCO® Human Skeletal Myoblasts respond to physiological levels of TGF-β1 and IGF-1. TGF-β1 addition inhibitsdifferentiation, whereas the addition of IGF-1 (Cat. No. PHG0075) blunts this effect. The anti-troponin staining method was used as described in Figure 1.

In the example above, Smad3 phosphorylation in response to TGF−β1 or myostatin was measured by TR-FRET after transduction of myotubes with BacMam-encoding LanthaScreen® Smad3-GFP.

Figure 3. Utilizing the BacMam delivery system, differentiated HSkM can be transduced with LanthaScreen® pathway sensors, enabling compound screeningwith respect to specific signaling pathways.

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