StemXVivo Endoderm Kit

R&D Systems™ StemXVivo™ Endoderm Kit (Catalog # SC019). Media supplements and growth factors, to differentiate human pluripotent stem cells into definitive endoderm under serum-free conditions.

• Protocol for the Definitive Endoderm Differentiation of BG01V Human Embryonic Stem Cells

Background Information

Definitive endoderm forms during gastrulation of the vertebrate embryo and gives rise to the lining of the gut, as well as multiple important organ systems including the lungs, liver, and pancreas. Embryonic stem (ES) cells are self-renewing cells that have the ability to differentiate into each of the three germ layers. The differentiation of ES cells into definitive endoderm is an essential first step for the transformation of these cells into downstream cell types, which can be useful in toxicological, developmental, and therapeutic studies.

Kit Description

The StemXVivo Endoderm Kit (Catalog # SC019) contains specially formulated media supplements and growth factors, which can be used to differentiate human pluripotent stem cells into definitive endoderm under serum-free conditions. The quantity of each component in this kit is sufficient to make 250 mL of media for differentiation. This is enough media for the differentiation of two 60 mm plates or one 24-well plate. This is estimated to be enough media for the differentiation of 2.8 x 10⁷ pluripotent cells. The cells that are differentiated using this kit can be expected to be at least 70% positive for the definitive endoderm marker, SOX17. In addition, these cells express minimal to non-existent levels of SOX7, a marker for extra-embryonic endoderm.

Kit Contents

• Endoderm Base Media Supplement
• FGF basic
• Activin A
• Wnt-3a

Definitive Endoderm Differentiated BG01V Human Embryonic Stem Cells. BG01V human embryonic stem cells were differentiated using the StemXVivo Endoderm Kit (Catalog # SC019) and stained with A) 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), or C) 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 counterstained with DAPI (blue). Endoderm differentiated cells were also stained with D) a PE-conjugated Mouse Anti-Human CXCR4 Monoclonal Antibody (Catalog # FAB173P; filled histogram) or a PE-conjugated Mouse IgG2B Isotype Control (Catalog # IC0041P; open histogram). High levels of SOX17 and low levels of SOX7 indicate endoderm differentiation.