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