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iCell® Skeletal Myoblasts: A Model for Interrogating and Modulating Glucose Uptake

iCell Skeletal Myoblasts

Figure 1: iCell Skeletal Myoblasts Provide an Easy-to-use Model for Human Skeletal Muscle.
iCell Skeletal Myoblasts adhere to the culture plate post-thaw and begin to form myotubes within 24 hours. By day 3 in culture, a high percentage of myogenin-positive nuclei are observed, and cells can be assayed for insulin-dependent glucose uptake.

Skeletal muscle plays a pivotal role in modulating circulating glucose levels, and the impact of muscle dysfunction on the onset, pathology, and treatment of type 2 diabetes is the subject of intense basic and applied research.

CDI’s iCell® Skeletal Myoblasts provide an excellent model for investigating the etiology, pathology, and potential treatments of type 2 diabetes while Promega’s Ultra-Glo Recombinant Luciferase based detection reagents offer easy-to-use, highly sensitive, and robust detection reagents for a variety of cellular processes.  In culture, iCell Skeletal Myoblasts fuse to form myotubes (Figure 1) that exhibit an approximately 2-fold increase in insulin-dependent glucose uptake as well as insulin-independent glucose uptake (Figure 2), and appropriate downstream phosphorylation events (Figure 3).  The recapitulation of native human glucose uptake mechanisms and simple handling of iCell Skeletal Myoblasts along with the non-radioactive robust detection and quantification of the Ultra-Glo reagents make this an ideal preparation for model development and screening in diabetes drug discovery efforts.

See the Protocol: Modulating Glucose Uptake in Skeletal Myotubes

Contact CDI’s Technical Support to learn more about glucose uptake and other ways CDI can further your research through iPSC technology.

Contact Promega’s Custom Assay Services  to learn more about the Ultra-Glo Recombinant Luciferase line of detection reagents and the Glucose Uptake Assay.

Contact Cisbio Bioassays to learn more about HTRF technology and their other assays for measuring cell signaling events.

References:

  1. Modulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis Cellular Dynamics Application Protocol
2-DG Uptake Assay

Figure 2: iCell Skeletal Myoblasts Provide a Robust Model for Insulin-dependent and Insulin-independent Glucose Uptake.
Stimulation with insulin (Ins) results in an approximately 2-fold increase in 2-DG uptake in a dose-dependent manner (inset). Treatment with the GLUT4-selective inhibitor, Indinavir, attenuates the assay signal and confirms the involvement of GLUT4 in this biological response. Insulin-independent glucose uptake is shown through induction with the beta-adrenergic agonists isoproterenol (Iso) and clenbuterol (Clen). All measurements were obtained via the Bioluminescent Glucose Uptake Assay Kit (Promega).

Figure 3: iCell Skeletal Myoblasts Provide a Complete Biological Model for Insulin-dependent Glucose Uptake.
The well-characterized AKT signaling pathway involved in insulin-dependent glucose uptake is also activated in iCell Skeletal Myoblasts with phosphorylation of S473 in a dose-dependent manner following insulin exposure (mean ± SD, n=3).