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).
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.
For Research Use Only. Not for use in diagnostic procedures.