cat no | io1002
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).
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.
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.
per vial
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@خقسا¶جتسئµ.
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.
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.
ioSkeletal Myocytes express skeletal myocyte-specific markers
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: 800آµm
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.
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
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; 50آµM 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.
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.
Starting material
Human iPSC line
Karyotype
Normal (46, XY)
Seeding compatibility
6, 12, 24, 48, 96 & 384 well plates
Shipping info
Dry ice
Donor
Caucasian adult male (skin fibroblast)
Vial size
Small: >2.5 x 10âپ¶ viable cells
Large: >5 x 10âپ¶ viable cells
Quality control
Sterility, protein expression (ICC) and gene expression (RT-qPCR)
Differentiation method
opti-ox deterministic cell programming
Recommended seeding density
100,000 cells/cm2
User storage
LN2 or -150آ°C
Format
Cryopreserved cells
Product use
ioCells are for research use only
Applications
Muscle and neuromuscular research
Amenable to high throughput screening
Contractility assays
3D co-cultures
Bernard, et al
خقسا¶جتسئµ
2024
خقسا¶جتسئµ
V5
خقسا¶جتسئµ
2023
Dr Marieke Aarts | Principal Scientist | Bi/ond
Amanda Turner | Senior Product Manager | خقسا¶جتسئµ
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.
Consistent. Defined. Scalable.
Dr Shushant Jain
Group Leader | In Vitro Biology | Charles River, 2021
Amy Rochford
PhD Neural Engineering and Bioelectronics | Cambridge University
Dr Michael Duchen
Professor of Physiology | University College London