cat no | io1002
ioSkeletal Myocytes
Human iPSC-derived
skeletal myocytes
ioSkeletal Myocytes, are human iPSC-derived skeletal myocytes deterministically programmed 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).
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Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
Benchtop benefits
Consistent
Batch-to-batch reproducibility and homogeneity create a stable human model for the study of muscle, neuromuscular, and associated metabolic disorders.
Quick
Form striated, multinucleated, myocytes by day 10 post revival, that contract in response to acetylcholine.
Easy-to-use
Cells arrive programmed to rapidly mature upon revival. One medium is required in a two-step protocol.
Ready within days
ioSkeletal Myocytes generated by transcription factor-driven deterministic programming of iPSCs using opti-ox technology
Time-lapse video capturing the rapid and homogeneous skeletal myocytes phenotype acquisition upon thawing of cryopreserved ioSkeletal Myocytes. 10 day time course.
Highly characterised and defined
ioSkeletal Myocytes express skeletal myocyte-specific markers
DAPI (blue)
DAPI (blue)
MHC (red)
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 deterministic programming
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
Click on the tabs to explore the data.
(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
Click on the tabs to explore the data.
(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.
Seeding density
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 mature rapidly upon revival in the recommended medium. The protocol for the generation of these cells is a two-phase process. Phase 1. Stabilisation for 3 days. Phase 2. Maintenance during which the skeletal myocytes mature.
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Cell culture hacks | human iPSC-derived skeletal myocytes
Read this blog on skeletal myocytes cell culture for our top tips on careful handling, cell plating and media changes to achieve success from the outset.
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What scientists say about ioSkeletal Myocytes
Dr Shushant Jain
Group Leader | In Vitro Biology | Charles River, 2021
“One of the biggest advantages of the ioSkeletal Myocytes is within the early drug discovery phase. You can very quickly screen a large number of molecules in a short amount of time with minimal variability and high reproducibility.”
Amy Rochford
PhD Neural Engineering and Bioelectronics | Cambridge University
"The ioSkeletal Myocytes have a much shorter cell culture time compared to harvesting primary muscle cells, saving us months on cell culture work. Another advantage of these cells is their higher population purity compared to other stem cell derived cells. This enables us to achieve higher numbers of functional striated muscle that are capable of contracting under electrical stimulation. "
Dr Michael Duchen
Professor of Physiology | University College London