Stem cell experiments require significant time, effort, and funding. The use of low quality or undefined reagents can introduce unwanted experimental variability that can compromise the validity of new data and raise doubts about previous conclusions. To increase the probability of experimental consistency and success, R&D Systems manufactures premium quality reagents for stem cell culture, characterization, expansion, and differentiation:
Defined Culture Matrix & Specialized Media
Highly defined reagents to reduce cell culture variability.
Premium Quality Cytokines & Growth Factors
Superior quality recombinant and natural proteins with exceptional lot-to-lot consistency, and high bioactivity to ensure consistent expansion & differentiation results.
GMP certified to reduce variability during the transition from pre-clinical to clinical research phases.
High Performance Antibodies
Over 12,000 antibodies to facilitate a thorough characterization of differentiation status & signal transduction mechanisms.
Specialized kits to verify the potency status of your starting cell population.
A complementary collection of reagents to investigate the signaling cascades that control stem cell proliferation and differentiation.
Definitive Endoderm Differentiation of iPS2 Human Induced Pluripotent Stem Cells. iPS2 Human Induced Pluripotent Stem Cells were cultured to 50% confluency in MEF Conditioned Media (Catalog # AR005) then differentiation to definitive endoderm was induced using 100 ng/mL Recombinant Human Activin A (Catalog # 338-AC) and 75 ng/mL Recombinant Human Wnt-3a (Catalog # 5036-WN). After 24 hours, the media was replaced with media containing 100 ng/mL of Recombinant Human Activin A alone and the cells were cultured for an additional 2 days. The cells were subsequently fixed with 4% PFA for 20 minutes and markers of definitive endoderm were simultaneously detected by immunocytochemistry. The nuclear protein SOX17 was detected using a Goat Anti-Human SOX17 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1924), and the cells were stained using the NorthernLights™ (NL) 493-conjugated Anti-Goat IgG Secondary Antibody (Catalog # NL003; green). The membrane-associated adhesion protein Claudin-6 was detected using a Mouse Anti-Human Claudin-6 Monoclonal Antibody (Catalog # MAB3656), and the cells were stained using the NL557-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # NL007; red). The nuclei were counterstained with DAPI (blue). Simultaneous detection of both SOX17 and Claudin-6 verified that successful definitive endoderm differentiation was induced by Activin A and Wnt-3a. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Recent Publications Citing R&D Systems Featured Products for Definitive Endoderm Differentiation
Wang, P. et al. (2012) A molecular signature for purified definitive endoderm guides differentiation and isolation of endoderm from mouse and human embryonic stem cells.
Stem Cells Dev. 21:2273.
Recombinant Human/Mouse/Rat Activin A (Catalog # 338-AC)
Recombinant Mouse Wnt-3a (Catalog # 1324-WN) Sample: Human and mouse embryonic stem cells Application: Bioassay