ioSkeletal Myocytes, are human iPSC-derived skeletal myocytes precision reprogrammed using opti-ox™ technology. Skeletal myocytes are delivered cryopreserved, upon revival the cells rapidly mature forming elongated striated, multinucleated muscle cells that contract within 10 days. Easy to culture, skeletal myocytes consistently exhibit high population purity expressing key myofilament proteins such as Desmin and Myosin Heavy Chain (MHC).
ioSkeletal Myocytes provide a source of highly-defined, consistent and reliable human muscle cells for research, disease modelling and high-throughput screening across areas such as muscle, neuromuscular, and associated metabolic disorders.
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D. Troponin (green) / Phalloidin (red) / DAPI (blue)
Immunocytochemistry staining at day 10 post revival demonstrates robust expression of components of the contractile apparatus such as Desmin (A), Dystrophin (B), and Myosin Heavy Chain (C), along with the muscle transcription factor Myogenin (C). Cells also demonstrate expression of Troponin with visible striated fibres and multinucleation (D).
Cells demonstrate classical myocyte morphology
ioSkeletal Myocytes form elongated multinucleated myocytes over 10 days. Day 1 to 10 post-thawing; 4X magnification; scale bar: 800µm
Cells demonstrate gene expression of key myogenic markers following reprogramming
Following reprogramming, ioSkeletal Myocytes downregulate expression of the pluripotency genes (A), whilst demonstrating robust expression of key myogenic markers (B). Gene expression levels assessed by RT-qPCR (data expressed relative to the parental hiPSC, normalised to HMBS). Data represents day 10 post-revival samples; n=7 biological replicates.
Robust and scalable cells for high-throughput screening
Cells are suitable for phenotypic based high-throughput screening
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(A) Immunocytochemistry | Human fibroblasts were transduced with lentiviral vectors allowing inducible over-expression of MYOD1 to transdifferentiate them to myocytes in approximately 10 days. Transdifferentiated myotubes were stained for multiple myotube markers to assess the purity and degree of multi-nucleation. (B) Immunocytochemistry | ioSkeletal Myocytes generate myocytes within as little as 4 days post-revival with a high-degree of MHC+ cells (>80% purity), suitable for phenotypic based high throughput screens. (C)Myosin Heavy Chain Positive Cells | The total area of MHC positive cells generated is similar in a comparison between ioSkeletal Myocytes and transdifferentiated fibroblasts.
Shushant Jain et al, Charles River Laboratories
Rapid gain of functionality
Cells express the insulin regulated glucose transporter GLUT4, critical for metabolic studies
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(A) Gene expression | RT-qPCR, at day 10 post-revival, demonstrating expression of GLUT4 in the ioSkeletal Myocytes, compared to undifferentiated human iPSCs and ioGlutamatergic Neurons.(B) Immunocytochemistry |at day 7 post-revival, ioSkeletal Myocytes express GLUT4 in peri-nuclear regions, and show striations. (C) Western blotting | analysis of differentiated 3T3-L1 adipocytes and maturing ioSkeletal Myocytes demonstrates GLUT4 expression in a time-dependent manner.
Dougall Norris & Daniel Fazakerley, Wellcome-MRC Institute of Metabolic Science
In vitro human muscle cells suitable for contractility assays
By day 10 post-revival, cells demonstrate a strong contractile response upon addition of acetylcholine, providing a suitable human muscle model for contractility assays. Spontaneous contraction is also observed during continuous culture (data not shown). Day 10 post-revival skeletal myocytes; 50µM acetylcholine.
Available in two vial sizes, tailored to suit your experimental needs with minimal waste
Recommended seeding density for ioSkeletal Myocytes is 100,000 cells/cm2.
One Small vial can plate a minimum of 0.5 x 24-well plate, 0.75 x 96-well plate, or 1 x 384-well plate.
One Large vial can plate a minimum of 1 x 24-well plate, 1.5 x 96-well plates, or 2 x 384-well plates.
Cells arrive ready to plate
ioSkeletal Myocytes are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended medium. The protocol for the generation of these cells is a three-phase process: Phase 0. Induction (carried out at bit.bio) Phase 1. Stabilisation for 3 days. Phase 2. Maintenance during which the skeletal myocytes mature.