STEMdiff™ Definitive Endoderm Differentiation Kit

总览

STEMdiff™ Definitive Endoderm Kit 是一套完整的无血清、无动物源成分的培养基与添加物剂盒，可高效支持人胚胎干细胞（hESC）和人诱导多能干细胞（hiPSC）向定向内胚层细胞分化。

使用 STEMdiff™ Definitive Endoderm Kit 分化的细胞高表达内胚层标志物，包括 CD184（CXCR4）、SOX17、FOXA2 和 c-KIT，同时不表达外胚层、中胚层及多能性标志物。通过该试剂盒获得的定向内胚层细胞具有多向分化潜能，可进一步分化为胰腺、肠道、肺及肝脏谱系细胞，因此是发育生物学研究、疾病建模及药物发现的强大工具。

本试剂盒针对在 mTeSR™1、mTeSR™ Plus 或 TeSR™-AOF 中维持培养的细胞的分化进行了优化。若需了解在 TeSR™-E8™ 中培养的细胞的分化，请参阅 STEMdiff™ Definitive Endoderm Kit (TeSR™-E8™ Optimized)。

分类
专用培养基

细胞类型
气道细胞，内胚层，PSC衍生，肝细胞，肠道细胞，胰腺细胞，多能干细胞

种属
人

应用
细胞培养，分化

品牌
STEMdiff

研究领域
癌症，上皮细胞研究，干细胞生物学

制剂类别
不含动物成分，无血清

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实验数据

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.

Density plots and quantitative analysis 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.

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.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

STEMdiff™ Definitive Endoderm Kit

Catalog #

05110

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

STEMdiff™ Definitive Endoderm Kit

Catalog #

05110

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

STEMdiff™ Definitive Endoderm Kit

Catalog #

05110

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

STEMdiff™ Definitive Endoderm Kit

Catalog #

05110

Lot #

All

Language

English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area

Workflow Stages

Workflow Stages for Human Pluripotent Stem Cell Research

Reprogramming

Maintenance

Differentiation

Workflow Stages for hPSC-Derived Endoderm Cell Research

Endoderm Induction

Characterization and Isolation

Downstream Differentiation

相关材料与文献

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.

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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.

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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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