STEMdiff™

STEMdiff™

Take the “If” Out of “Diff”

Human pluripotent stem cell (hPSC)-based models hold tremendous potential for the study of human development and disease. When working with hPSCs, even the most detailed and rigorously-followed protocols can still lead to inconsistent differentiation.1,2 Use STEMdiff™ to reproducibly differentiate across multiple human embryonic stem (ES) cell and induced pluripotent stem (iPS) cell lines. STEMdiff™ is part of the most complete system of reagents for human pluripotent stem cell (hPSC) culture - a system that includes mTeSR™1, the most published feeder-free ES cell and iPS cell culture medium. Media formulations, raw material specifications and manufacturing processes for all STEMdiff™ products have been carefully optimized in order to minimize reagent variability. In addition, STEMdiff™ products include detailed, user-friendly protocols to standardize your differentiation procedures.

Why Use STEMdiff™?

  • Reduce variability between experiments.
  • Use with hPSCs cultured with TeSR™ maintenance media.
  • Optimized formulations and pre-screened components ensure reagentconsistency and minimal lot-to-lot variability.
  • Detailed protocols for differentiating to all three germ layers.
  • Efficient differentiation of multiple human ES and iPS cell lines.

Which Lineages Are You Interested in Differentiating to?

Ectodermal Lineages

Protocol Type:

Features/Advantages:

  • Rapid neural induction using EB or monolayer protocols
  • Serum-free* expansion and cryopreservation of neural progenitor cells
  • Highly efficient generation of neuronal and glial precursors from neural progenitor cells
  • Maturation of precursors to functional forebrain-type neurons, dopaminergic neurons or astrocytes
*STEMdiff™ Astrocyte Differentiation Kit contains serum.

Mesodermal Lineages

Protocol Type:

Monolayer

Features/Advantages:

  • Rapid mesodermal induction after only 2 - 4 days of differentiation
  • Generate multipotent early mesoderm cells that can be further differentiated to multiple downstream mesodermal cell lineages including endothelial, osteogenic, chondrocytes and adipocytes

Endodermal Lineages

Protocol Type:

Monolayer

Features/Advantages:

  • Generate multipotent definitive endoderm cells that can be differentiated to multiple downstream endodermal cell lineages including hepatic, pancreatic, intestinal and pulmonary
  • Generate functional pancreatic progenitor cells that can be further differentiated to insulin-producing β-cells

Flexible User-Directed Differentiation

Protocol Type:

Embryoid body (EB) or monolayer

Features/Advantages:

  • Unsupplemented medium without supports cell survival without lineage bias
  • Can be used to direct differentiation to a variety of cell lineages, including endothelial,3 smooth muscle,3 hematopoietic,4,5 skeletal muscle,6 and cardiomyocytes7
  • Allow customization of differentiation protocols
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STEMdiff™ Neural System for hPSC-Based Neurological Modeling

hPSC-derived neural cells provide a physiologically relevant model for drug discovery, cell therapy validation and neurological disease research. Watch this video to discover how you can use the STEMdiff™ Neural System to generate, expand, differentiate, characterize and cryopreserve your hPSC-derived neural progenitor cells.

AggreWell™: For Uniform Embryoid Body Formation

Many protocols for differentiating human ES and iPS cells start with the formation of embryoid bodies (EBs). Conventional EB formation methods result in EBs of heterogeneous size and shape, leading to inefficient and uncontrolled differentiation.8,9 AggreWell™ plates allow users to produce uniformly shaped EBs of controlled size, making differentiation experiments more reproducible. 10

References

  1. D’Amour KA et al. (2005) Efficient differentiation of human embryonic stem cells to definitive endoderm. Nature Biotechnology 23(12):1534-41
  2. Kattman SJ et al. (2011) Stage-Specific Optimization of Activin/Nodal and BMP Signaling Promotes Cardiac Differentiation of Mouse and Human Pluripotent Stem Cell Lines. Cell Stem Cell 8(2): 228-240
  3. Tan JY et al. (2013) Efficient Derivation of Lateral Plate and Paraxial Mesoderm Subtypes from Human Embryonic Stem Cells Through GSKi-Mediated Differentiation. Stem Cells Dev 22(13): 1893-1906
  4. Protocol adapted from Chadwick et al. (2003) and Ng et al. (2008), and discussed in this Technical Bulletin
  5. Gertow et al. (2013) WNT3A Promotes Hematopoietic or Mesenchymal Differentiation from hESCs Depending on the Time of Exposure. Stem Cell Reports 1(1):53-65
  6. Xu C et al. (2013) A Zebrafish Embryo Culture System Defines Factors that Promote Vertebrate Myogenesis across Species. 155(4): 909-921
  7. Elliott DA et al. (2011) NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes. Nat Methods 8(12):1037-1040
  8. Bauwens CL et al. (2008) Control of human embryonic stem cell colony and aggregate size heterogeneity influences differentiation trajectories.Stem Cells 26(9): 2300-2310
  9. Bratt-Leal AM, et al. (2009) Engineering the embryoid body microenvironment to direct embryonic stem cell differentiation. Biotechnol Prog 25(1): 43-51
  10. Yanai A, et al. (2013) Differentiation of Human Embryonic Stem Cells Using Size-Controlled Embryoid Bodies and Negative Cell Selection in the Production of Photoreceptor Precursor Cells. Tissue Eng Part C Methods 19(10): 755-764
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