STEMdiff™ Myogenic Progenitor Supplement Kit

Serum-free supplements for differentiation of human pluripotent stem cells (hPSCs) to myogenic progenitor cells
STEMdiff™ Myogenic Progenitor Supplement Kit

Serum-free supplements for differentiation of human pluripotent stem cells (hPSCs) to myogenic progenitor cells

1 Kit
Catalog # 100-0151
579 USD
Required Products
  1. mTeSR™1
    mTeSR™1

    cGMP, feeder-free maintenance medium for human ES and iPS cells

  2. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  3. D-PBS Without Ca++ and Mg++
    D-PBS (Without Ca++ and Mg++)

    Dulbecco’s phosphate-buffered saline without calcium and magnesium

  4. Collagenase Type IV (1 mg/mL)
    Collagenase Type IV (1 mg/mL)

    Cell dissociation reagent

Overview

STEMdiff™ Myogenic Progenitor Supplement Kit comprises serum-free supplements for the differentiation of human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells to myogenic progenitor cells. These cells, which are characterized by myogenic cell markers such as CD56 and CD82, can be culture-expanded for 5+ passages using MyoCult™-SF Expansion Supplement Kit (Human; Catalog #05980) and further differentiated to functional multinucleated MyHC+ myotubes at high efficiency using MyoCult™ Differentiation Kit (Human; Catalog #05965). These myotubes can be used in various downstream applications and analyses. This kit is compatible with human ES and iPS cells maintained in mTeSR™ 1 (Catalog #85850), mTeSR™ Plus (Catalog #100-0276), or TeSR™-E8™ (Catalog #05990).
Advantages
⦁ Efficient differentiation to myogenic pregenitors across human PSC lines
⦁ PSC-derived myotubes display organized sarcomeric proteins and contractility
⦁ Optimized reagents designed to support the entire human PSC-derived myogenic progenitor workflow
Components
  • STEMdiff™ Myogenic Progenitor Supplement A, 240 μL
  • STEMdiff™ Myogenic Progenitor Supplement B, 240 μL
  • STEMdiff™ Myogenic Progenitor Supplement C, 240 μL
  • STEMdiff™ Myogenic Progenitor Supplement D, 60 mL
Subtype
Supplements
Cell Type
Myogenic Stem and Progenitor Cells
Species
Human
Application
Cell Culture, Differentiation
Brand
STEMdiff
Area of Interest
Disease Modeling, Stem Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
STEMdiff™ Myogenic Progenitor Supplement Kit
Catalog #
100-0151
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™ Myogenic Progenitor Supplement Kit
Catalog #
100-0151
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™ Myogenic Progenitor Supplement Kit
Catalog #
100-0151
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
STEMdiff™ Myogenic Progenitor Supplement Kit
Catalog #
100-0151
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
STEMdiff™ Myogenic Progenitor Supplement Kit
Catalog #
100-0151
Lot #
All
Language
English

Educational Materials (5)

Brochure
Culture and Expand Mouse Myogenic Progenitors
Brochure
mTeSR™ Plus Product Overview Flyer
Wallchart
Derivation and Applications of Human Pluripotent Stem Cells
Wallchart
SnapShot: Exercise Metabolism
Webinar
3D Modeling of Skeletal Muscle for In Vitro Functional Assaying
55:57
3D Modeling of Skeletal Muscle for In Vitro Functional Assaying

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Cell images represinting different stages of myogenic differentiation use STEMdiff Myogenic Progenirot Supplement kit.

Figure 1. Schematic of Myogenic Induction Using the STEMdiff™ Myogenic Progenitor Supplement Kit

Prior to differentiation (Day -1), human pluripotent stem cells (hPSCs) are plated in Corning® Matrigel®-coated plates. On Day 0, differentiation is initiated and continued by performing daily media changes with STEMdiff™ Myogenic Progenitor Supplement A at Days 0 - 2, STEMdiff™ Myogenic Progenitor Supplement B at Days 2 - 4, STEMdiff™ Myogenic Progenitor Supplement C at Days 4 - 6, and STEMdiff™ Myogenic Progenitor Supplement D for the remainder of the protocol (Days 6 - 30). Upon completion of the 30-day protocol, the culture contains an abundance of PAX7+MyoD+ myogenic progenitors that can be further expanded using MyoCult™-SF Expansion Supplement Kit (Human; Catalog #05980) for use in downstream applications.

Heatmap of differential gene expression for myogenic progenitors differentiated from embryonic stage.

Figure 2. hPSCs Differentiated Using STEMdiff™ Myogenic Progenitor Kit Demonstrate Transcript Patterns of Embryonic Muscle Development

hPSCs (H9 cell line) were differentiated using the STEMdiff™ Myogenic Progenitor Kit and harvested at indicated time points for transcript analysis via qPCR; transcription was normalized against 18s as the housekeeping gene. The resulting heatmap indicates transcriptional changes to genes that mark specific stages of muscle development (mesoderm, paraxial mesoderm, somites, and myogenesis) and demonstrates a cellular trajectory that resembles embryonic muscle development.

Flow cytometry analysis of myogenic progenitors generated using the STEMdiff™ Myogenic Progenitor Kit.

Figure 3. STEMdiff™ Myogenic Progenitor Kit Generates Expandable hPSC-Derived Myogenic Progenitors

(A) Representative image of proliferating sub-cultured hPSC-derived myogenic progenitors generated using the STEMdiff™ Myogenic Progenitor Kit. (B) hPSC-derived myogenic progenitors harvested at passage 1 and 5 expressed human myoblast markers CD56 and CD82. (C) Expansion rates of hPSC-derived myogenic progenitors (hSPC-MP) over 5 passages across multiple hPSC lines are comparable to human primary myoblasts. Error bars represent standard error of mean, n = 3.

Microscopy images of different hPSC cell lines differentiated to myogenic progenitors using STEMdiff™  Myogenic Progenitor Kit and stained for myogenic markers MgHC and MYOD.

Figure 4. STEMdiff™ Myogenic Progenitor Kit Facilitates Efficient Differentiation of hPSC-Derived Myogenic Progenitors into hPSC-Derived Myotubes

(A) hPSC-derived myogenic progenitors were generated from multiple hPSC lines using the STEMdiff™ Myogenic Progenitor Kit and then induced to differentiate into myotubes using MyoCult™ Differentiation Medium (Human; Catalog #05965). After 8 days, myotubes were fixed and stained for MyHC and MyoD. (B) Multiple hPSC cell lines differentiated and induced using said method exhibited high fusion indices similar to primary human myoblasts (Numbers in bars represent the n number and dots represent technical replicates).

Microscopy image and electrophysiology of myotubes differentiated from hPSC-generated myogenic progenitors using STEMdiff™ Myogenic Progenitor Kit.

Figure 5. hPSC-Derived Myotubes Generated Using STEMdiff™ Myogenic Progenitor Kit Are Functionally Contractile

hPSC-derived myogenic progenitors were generated using the STEMdiff™ Myogenic Progenitor Kit and then induced to differentiate into myotubes using MyoCult™ Differentiation Medium. (A) hPSC-derived myotubes were stained for alpha-actinin and displayed organized sarcomeric structures as indicated by the zoomed-in area. (B) Spontaneous field potential recordings of hPSC-derived myotubes using microelectrode assay plate indicated that the derived myotubes are contractile. (C) Representative video of contracting hPSC-derived myotubes.

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