Defined, serum-free, animal component-free medium for the differentiation of human ES and iPS cells to definitive endoderm in a complete, ready-to-use format
Efficient and reproducible differentiation of multiple ES cell and iPS cell lines
Generates definitive endoderm cells capable of further differentiation to pancreatic, hepatic, intestinal and pulmonary cell lineages
产品组分包括
STEMdiff™ Endoderm Basal Medium, 100 mL
STEMdiff™ Definitive Endoderm Supplement MR (100X), 0.35 mL
STEMdiff™ Definitive Endoderm Supplement CJ (100X), 1.1 mL
STEMdiff™ Definitive Endoderm Kit is a complete, serum- and animal component-free medium and supplement kit that supports highly efficient differentiation of human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells to definitive endoderm cells. Cells differentiated using STEMdiff™ Definitive Endoderm Kit express high levels of endoderm markers, including CD184 (CXCR4), SOX17, FOXA2, and c-KIT, and lack expression of ectoderm, mesoderm, and pluripotency markers. The definitive endoderm cells produced using this kit are multipotent and capable of further differentiation towards cells of the pancreatic, intestinal, pulmonary, and hepatic lineages, thus providing a robust tool for developmental studies, disease modeling, and drug discovery.
Figure 1. Definitive endoderm differentiation is efficient across multiple human ES and iPS cell lines
Quantitative analysis of definitive endoderm formulation on multiple human ES and iPS cell lines as measured by co-expression of CXCR4 and SOX17. Prior to differentiation using STEMdiff™ Definitive Endoderm, cells were maintained in their pluripotent state by culturing mTeSR™1 on Matrigel. Data are expressed as the mean percent of cells expressing both markers. Error bars indicate SEM, n = 4-18 per cell line.
Figure 2. Quantitative Analysis of Definitive Endoderm hES and iPS-Derived Using STEMdiff™ Definitive Endoderm
Quantitative analysis of definitive endoderm in human ES and iPS cells previously maintained in TeSR™2 prior to differentiation on Matrigel using STEMdiff™ Definitive Endoderm. Data are expressed as the mean percent of cells expressing both markers. Error bars indicate SEM. n = 4-11 per cell line.
Figure 3. Efficient definitive endoderm differentiation in human ES and iPS cells
Representative Density plots showing CXCR4 and SOX17 expression in human ES cells (H1 and H9) and human iPS cells (WLS-4D1 and A13700) following 5 days of differentiation to definitive endoderm using STEMdiff™ Definitive Endoderm. Isotype controls were used to set quadrant gates.
Figure 4. STEMdiff™ Definitive Endoderm yields DE that retains potency for downstream lineage specification
Cultures differentiated using STEMdiff™ Definitive Endoderm maintain their ability to be directed towards pancreatic and hepatic lineages. A) Representative image of PDX-1 immunoreactivity in H9 cells following pancreatic specification. Scale bar 20 µm. B) Representative image of human serum albumin (HSA) immunoreactivity in H9 cells following hepatic specification. Scale bar, 100 µm.
Figure 5. Generation of Definitive Endoderm from hPSCs Maintained in mTeSR™ Plus
(A) Representative density plots showing CXCR4 and SOX17 expression in cells cultured in mTeSR™1 (daily feeds) or mTeSR™ Plus (restricted feeds), following 5 days of differentiation using the STEMdiff™ Definitive Endoderm Kit. (B) Quantitative analysis of definitive endoderm formation in multiple hPSC lines (H9, STiPS-M001, WLS-1C) maintained with mTeSR™1 or mTeSR™ Plus as measured by co-expression of CXCR4 and SOX17. Data are expressed as the mean percentage of cells (± SEM) expressing both markers; n=3.
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Comparative characterization of human induced pluripotent stem cells (hiPSC) derived from patients with schizophrenia and autism. L.-M. Grunwald et al. Translational psychiatry 2019
Abstract
Human induced pluripotent stem cells (hiPSC) provide an attractive tool to study disease mechanisms of neurodevelopmental disorders such as schizophrenia. A pertinent problem is the development of hiPSC-based assays to discriminate schizophrenia (SZ) from autism spectrum disorder (ASD) models. Healthy control individuals as well as patients with SZ and ASD were examined by a panel of diagnostic tests. Subsequently, skin biopsies were taken for the generation, differentiation, and testing of hiPSC-derived neurons from all individuals. SZ and ASD neurons share a reduced capacity for cortical differentiation as shown by quantitative analysis of the synaptic marker PSD95 and neurite outgrowth. By contrast, pattern analysis of calcium signals turned out to discriminate among healthy control, schizophrenia, and autism samples. Schizophrenia neurons displayed decreased peak frequency accompanied by increased peak areas, while autism neurons showed a slight decrease in peak amplitudes. For further analysis of the schizophrenia phenotype, transcriptome analyses revealed a clear discrimination among schizophrenia, autism, and healthy controls based on differentially expressed genes. However, considerable differences were still evident among schizophrenia patients under inspection. For one individual with schizophrenia, expression analysis revealed deregulation of genes associated with the major histocompatibility complex class II (MHC class II) presentation pathway. Interestingly, antipsychotic treatment of healthy control neurons also increased MHC class II expression. In conclusion, transcriptome analysis combined with pattern analysis of calcium signals appeared as a tool to discriminate between SZ and ASD phenotypes in vitro.
Intrinsic Immunity Shapes Viral Resistance of Stem Cells. Wu X et al. Cell 2018 JAN
Abstract
Stem cells are highly resistant to viral infection compared to their differentiated progeny; however, the mechanism is mysterious. Here, we analyzed gene expression in mammalian stem cells and cells at various stages of differentiation. We find that, conserved across species, stem cells express a subset of genes previously classified as interferon (IFN) stimulated genes (ISGs) but that expression is intrinsic, as stem cells are refractory to interferon. This intrinsic ISG expression varies in a cell-type-specific manner, and many ISGs decrease upon differentiation, at which time cells become IFN responsive, allowing induction of a broad spectrum of ISGs by IFN signaling. Importantly, we show that intrinsically expressed ISGs protect stem cells against viral infection. We demonstrate the in vivo importance of intrinsic ISG expression for protecting stem cells and their differentiation potential during viral infection. These findings have intriguing implications for understanding stem cell biology and the evolution of pathogen resistance.
Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen Carpentier A et al. Stem Cell Research 2016 MAR
Abstract
The establishment of protocols to differentiate human pluripotent stem cells (hPSCs) including embryonic (ESC) and induced pluripotent (iPSC) stem cells into functional hepatocyte-like cells (HLCs) creates new opportunities to study liver metabolism, genetic diseases and infection of hepatotropic viruses (hepatitis B and C viruses) in the context of specific genetic background. While supporting efficient differentiation to HLCs, the published protocols are limited in terms of differentiation into fully mature hepatocytes and in a smaller-well format. This limitation handicaps the application of these cells to high-throughput assays. Here we describe a protocol allowing efficient and consistent hepatic differentiation of hPSCs in 384-well plates into functional hepatocyte-like cells, which remain differentiated for more than 3 weeks. This protocol affords the unique opportunity to miniaturize the hPSC-based differentiation technology and facilitates screening for molecules in modulating liver differentiation, metabolism, genetic network, and response to infection or other external stimuli.
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