StemXVivo Endoderm Kit
R&D Systems, part of Bio-Techne | Catalog # SC019B
For endoderm differentiation of human pluripotent stem cells
Key Product Details
Species
Human
Product Summary for StemXVivo Endoderm Kit
Kit Summary
A kit optimized to efficiently differentiate pluripotent cells into definitive endodermal cells under serum-free conditions.
Key Benefits
- Definitive endoderm differentiation in 4 days
- Pre-optimized to reduce experimental variation
- Endodermal cells are 70–90% positive for SOX17
- Endodermal cells express minimal to non-detectable levels of SOX7
Why Use Pre-optimized Reagents for Endoderm Differentiation?
Pluripotent stem cells can differentiate into all three germ layers including the definitive endoderm which gives rise to cells of the liver, lung, pancreas, and gut lining. Inefficient or inconsistent differentiation represents a costly and time-consuming obstacle in stem cell differentiation.
High quality media and defined differentiation supplements can reduce experimental variability and increase differentiation efficiency.
The StemXVivo® Endoderm Kit:
- Includes premium quality reagents that are optimized to induce endoderm differentiation in 4 days.
- Includes components with low lot-to-lot variation to reduce experimental variability.
- Yields definitive endodermal cells expected to be at 70–90% positive for SOX17.
- Yields an endodermal cell population with little to non-detectable SOX7 expression.
Kit Contents
The StemXVivo® Endoderm Kit includes a serum-free, defined media supplement and premium quality recombinant proteins that have been optimized to efficiently differentiate pluripotent stem cells into definitive endodermal cells.
Cells are differentiated using two differentiation media that are made by combining the media supplement and the recombinant proteins included in this kit with RPMI medium. Differentiation Media I contains Activin A, FGF basic, and Wnt-3a whereas Differentiation Media II contains Activin A and FGF basic.
- Endoderm Base Media Supplement (50x)
- Recombinant Human Activin A
- Recombinant Human FGF Basic
- Recombinant Human Wnt-3a
- Endoderm Marker: Goat Anti-Human SOX17 Antigen Affinity-purified Polyclonal Antibody
Scientific Data Images for StemXVivo Endoderm Kit
Definitive Endoderm Differentiation of BG01V Human Embryonic Stem Cells.
BG01V cells were differentiated into definitive endoderm using the media supplements included in this kit (Catalog # SC019B). To evaluate lineage commitment, the cells were stained with the Anti-Human SOX17 antibody included in this kit. The cells were stained with NorthernLights™557-Conjugated Donkey Anti-Goat IgG Secondary Antibody (R&D Systems, Catalog # NL001; red) and the nuclei were counterstained with DAPI (blue).
The Differentiation of BG01V Human Embryonic Stem Cells into Definitive Endoderm Confirmed by Cell Markers (SOX17+/CXCR4+/FOX2A+/SOX7low).
BG01V human embryonic stem cells were differentiated using the StemXVivo®Endoderm Kit (Catalog # SC019) and stained withA) a Goat Anti-Human SOX17 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1924),B) a Goat Anti-Human HNF-3 beta/FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400), orC) a Goat Anti-Human SOX7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2766). In all images, cells were stained with the NorthernLights™557-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # NL001, red) and the nuclei were counterstained with DAPI (blue). Endoderm-differentiated cells were also stained withD) a PE-conjugated Mouse Anti-Human CXCR4 Monoclonal Antibody (Catalog # FAB173P; filled histogram) or a PE-conjugated Mouse IgG2BIsotype Control (Catalog # IC0041P; open histogram). High levels of SOX17, CXCR4, and HNF-3 beta/FoxA2 combined with low levels of SOX7 indicate definitive endoderm differentiation.
Differentiation of Human Amniotic Fluid Stem Cells into Definitive Endoderm.
The StemXVivo®Endoderm Kit (Catalog # SC019) was used to differentiate first-trimester human amniotic fluid stem cells (AFSCs) into definitive endoderm by culturing AFSCs in media containing recombinant human FGF basic, recombinant human Wnt-3a, and recombinant human Activin A for one day. On day 2, the media was removed and replaced with fresh media containing recombinant human FGF basic and recombinant human Activin A. The media was replaced twice daily until day 5 when cells were analyzed for expression of definitive endoderm markers.A.Before differentiation (day 0) AFSCs express the extra-embryonic endoderm marker SOX7, but not the definitive endoderm markers SOX17, HNF-3 beta, CXCR4, and GSC, as determined by quantitative real-time RT-PCR. After differentiation (day 5) into definitive endoderm, expression of SOX17, HNF-3 beta, CXCR4, and GSC was detected, but not expression of SOX7.***P< 0.001.B.Before differentiation (day 0) AFSCs fail to express the definitive endoderm marker CXCR4 as assessed by flow cytometry. However, after differentiation (day 5) the cells express CXCR4 (isotype control in black).***P< 0.001.Adapted from: Moschidou, D. et al. (2012) Mol. Therapy 20: 1953-1967
Formulation, Preparation, and Storage
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Background: Pluripotent Stem Cells
Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of pre-implantation embryos. Induced pluripotent stem (iPS) cells can be generated by somatic cell reprogramming following the exogenous expression of specific transcription factors (Oct-3/4, KLF4, SOX2, and c-Myc). These cell types are capable of unlimited, undifferentiated proliferation in vitro and still maintain the capacity to differentiate into a wide variety of somatic cells. In this capacity, pluripotent stem cells have widespread clinical potential for the treatments of heart disease, diabetes, spinal cord injury, and a variety of neurodegenerative disorders.
R&D Systems offers a wide range of products to support pluripotent stem cell culture and differentiation. Mouse embryonic fibroblasts may be used to maintain and expand pluripotent stem cells in an undifferentiated state. We also offer defined culture media, which are specifically optimized for use with human or rodent pluripotent stem cells. In addition, R&D Systems offers a variety of products to assess differentiation status and identify specific stem cell types of interest, including panels of marker antibodies, primer pairs, multi-color flow cytometry kits, and specialized verification kits.
Additional Pluripotent Stem Cells Products
Product Documents for StemXVivo Endoderm Kit
Product Specific Notices for StemXVivo Endoderm Kit
For research use only
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