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R&D SC019B StemXVivo Endoderm Kit
产品编号 SC019B| rndsystems
产品介绍
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. See Details
Data Examples
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 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 the nuclei were 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, 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
Expression of SOx17 and HNF-3 beta in Human Amniotic Fluid Stem Cells Differentiated into Definitive Endoderm.The StemXVivoEndoderm 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. Prior to differentiation (day 0), AFSCs show little to no expression of the definitive endoderm markers SOX17 and HNF-3 beta as assessed by confocal microscopy. However, after differentiation (Day 5), cells express both SOX17 and HNF-3 beta. The nuclei were stained with DAPI (blue). Scale Bar, 50 um.Adapted from: Moschidou, D. et al. (2012) Mol. Therapy 20: 1953-1967
BG01V human embryonic stem cells are licensed from ViaCyte, Inc.
Preparation and Storage
Stability & 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 vitroand 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.
公司简介
R&D Systems作为全球免疫学和细胞生物学产品的领跑者，1976年创立于美国明尼苏达州。其母公司Bio-Techne（原Techne公司）于1983年在美国NASDAQ上市。
目前，R&D Systems拥有近30,000种产品，95%以上由自己研发生产。其产品包括细胞因子、ELISA试剂盒、生长因子、趋化因子、抗体、Animal-Free™蛋白、Luminex 液相芯片、多因子检测固相芯片、流式细胞分析与细胞筛选、干细胞及细胞培养等；覆盖肿瘤、发育、糖生物、内分泌、免疫、神经学、蛋白酶、信号传导等学科研究领域；且每年有近2,000种新产品不断问世，以满足不同科研工作者的研究需要。
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