Batch to batch reproducibility and homogeneity create a stable human model for the study of muscle, neuromuscular, and associated metabolic disorders.
Human iPSC-derived skeletal myocytes
io1002 | Formerly known as ioMYOCYTES/skeletal, cat no: ea1200
Introducing 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. ioSkeletal Myocytes, part of our ioCells portofolio, have been reprogrammed from human induced pluripotent stem cells (iPSCs) using our precise reprogramming technology: opti-ox™¹.
The technology allows for human pluripotent stem cells, within days, to convert into skeletal myocytes, providing a high-quality, easy to use, cellular model.
Skeletal myocytes demonstrate robust expression of components of the contractile apparatus and form striated, multinucleated, myocytes by Day 10 post revival, that contract in response to acetylcholine.
Unprecedented batch-to-batch consistency allows for their use in demanding screening applications where small changes need to be detected against background noise.
Advantages
– Ready for experimentation within days
– Highly characterised and defined
– Easy culturing
Applications
– Muscle and neuromuscular research
– Amenable to high throughput screening
– Contractility assays
– 3D cocultures
– Single cell CRISPR Screening
For more information download our flyer
or email info@bit.bio
Batch to batch reproducibility and homogeneity create a stable human model for the study of muscle, neuromuscular, and associated metabolic disorders.
Form striated, multinucleated, myocytes by Day 10 post revival, that contract in response to acetylcholine.
Industrial scale quantities at a price point that allows the cells to be used from research to screening scale.
Cells arrive programmed to rapidly mature upon revival. One medium required in a two-step protocol.
ioSkeletal Myocytes express skeletal myocyte-specific markers
A. Desmin (red) / DAPI (blue)
B. Dystrophin (green) / DAPI (blue)
C. Myogenin (green) / MHC (red)
D. Troponin (green) / Phalloidin (red) / DAPI (blue)
Cells demonstrate classical myocyte morphology
Cells demonstrate gene expression of key myogenic markers following reprogramming
Cells express the insulin regulated glucose transporter GLUT4, critical for metabolic studies
(A) RT-qPCR, at Day 10 post-revival, demonstrating expression of GLUT4 in the ioSkeletal Myocytes, compared to undifferentiated hiPSCs and ioGlutamatergic Neurons (formerly known as ioNEURONS/glut). (B) Immunocytochemistry, at Day 7 post-revival, demonstrates expression of GLUT4 in peri-nuclear regions, and striations, in the ioSkeletal Myocytes*. (C) Western blotting 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
Cells are suitable for phenotypic based high-throughput screening
(A) 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) 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) Comparable total area of MHC positive cells are generated between ioSkeletal Myocytes and transdifferentiated fibroblasts.
Shushant Jain et al, Charles River Laboratories
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.
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.
Available in two vial sizes, tailored to suit your experimental needs with minimal waste
“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.”
Dr Shushant Jain
Group Leader | In Vitro Biology | Charles River
"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. This considerably increases the pace at which we can test our bioelectronics devices."
Amy Rochford
PhD Neural Engineering and Bioelectronics | Cambridge University
“We can now start asking questions that, ten years ago, we didn’t know how to answer,” Duchen reflects. “If you have a really good disease model, then the only limit is your imagination.”
Dr Michael Duchen
Professor of Physiology | University College London
Starting material
Human iPSC line
Donor
Caucasian adult male
(skin fibroblast)
Differentiation method
opti-ox™ cellular reprogramming
Karyotype
Normal (46, XY)
Vial size
Small: >2.5 x 106 viable cells
Large: >5 x 106 viable cells
Recommended seeding density
100,000 cells/cm2
Seeding compatibility
6 to 384 well plates
Quality control
Sterility, protein expression (IF) and gene expression (RT-qPCR)
User storage
LN2 or -150°C
Shipping info
Dry ice
Product use
These cells are for research use only