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STEMdiff™ Kidney Organoid Kit

Serum-free medium kit for the culture of kidney organoids from hPSCs
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产品优势


  • RELEVANT. Enables generation of human kidney organoids that model the developing nephron and associated endothelium and mesenchyme.

  • SIMPLE. Minimizes culture manipulations with a two-stage culture system and easy-to-follow protocol.

  • RELIABLE. Provides low experimental variability through an optimized formulation and rigorous quality controls.

  • HIGH THROUGHPUT. Reproducibly generates organoids in 96- and 384-well formats.

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    概述
    实验数据
    产品说明书及文档
    应用领域
    相关材料与文献
    STEMdiff™ Kidney Organoid Kit is a complete, serum-free cell culture medium system that supports highly efficient and reproducible generation of human pluripotent stem cell (hPSC)-derived kidney organoids in a simple, two-stage differentiation protocol. These kidney organoids are composed of podocytes, proximal tubules, distal tubules, as well as the associated endothelium and mesenchyme.

    Kidney organoids generated with STEMdiff™ Kidney Organoid Kit are tested for compatibility with phenotypic high-throughput assays such as nephrotoxic compound screening. They also provide a relevant and convenient model for studies related to developmental biology and disease modeling.

    The kit has been optimized for the differentiation of hPSCs previously cultured in mTeSR1 (Catalog #85850). It is also compatible with mTeSR Plus (Catalog #100-0276) maintained hPSCs, provided that mTeSR1 medium is used for the first three days of the protocol.

    Data Figures

    Figure 1. Representative Images of Kidney Organoids

    The STEMdiff™ Kidney Organoid Kit facilitates the directed differentiation of hPSCs to form kidney organoids that model the developing nephron. Pictured are human kidney organoids grown using the STEMdiff™ Kidney Organoid Kit differentiated from (A) iPS (WLS-1C) or (B) ES (H9) cells and imaged on day 12 and day 18 of differentiation, respectively.

    Figure 2. Schematic for Differentiation from hPSCs to Human Kidney Organoids with the STEMdiff™ Kidney Organoid Kit

    hPSC cultures progress through a simple two-stage process to generate kidney organoids. hPSCs are first plated and overlaid with Corning® Matrigel® to form cavitated spheroids. On the following day (Day 0), differentiation of cavitated PSC spheroids is initiated by changing medium from mTeSR™1 to STEMdiff™ Kidney Organoid Kit. During the next 18 days, the cells are directed through stages of the late primitive streak, intermediate mesoderm, and metanephric mesoderm to give rise to kidney organoids that are composed of podocytes, proximal and distal tubules, and an associated endothelium and mesenchyme.

    Figure 3. Efficient Differentiation of Human Pluripotent Stem Cells into Self-Organizing Kidney Organoids

    The STEMdiff™ Kidney Organoid Kit enables high-efficiency generation of kidney organoids from both ES (A, H1; B, H9) and iPS (C, WLS-1C; D, STiPS-M001) cell lines. (E) Kidney organoids were grown using the STEMdiff™ Kidney Organoid Kit or using home-made medium, and the average number of organoids per well of a 96-well plate was quantified on Day 18. All four tested cell lines were capable of differentiating into self-organizing kidney organoids that form convoluted tubular structures with high efficiency (mean ± SD, n ≥ 2).

    Figure 4. Kidney Organoids Grown Using the STEMdiff™ Kidney Organoid Kit Show the Expected Changes in Gene Expression During Differentiation

    As organoids progress through the stages of differentiation to more mature renal cell types, gene expression shifts from markers of pluripotency (day 0) to the mesoderm (day 1.5 - 4) and to the intermediate mesoderm/metanephric mesenchyme by day 4 - 12. Markers of the podocytes, proximal and distal tubules, mesenchyme and endothelium are observable starting by day 14. Marker levels were assessed in four independent experiments by RT-qPCR and normalized to expression levels of undifferentiated cells.

    Figure 5. Kidney Organoids form Convoluted Tubular Structures with Typical Nephron-like Segmentation

    (A) During differentiation, kidney organoids form convoluted tubular structures that resemble the structure and segmentation of the developing nephron. (B) These organoids express markers of the renal epithelium including podocyte marker podocalyxin (red), proximal tubule marker lotus tetragonolobus lectin (green), and distal tubule marker E-cadherin (white). Cultures also contain (C) endothelial cells stained by platelet endothelial cell adhesion molecule (CD31, white), as well as (D) mesenchymal markers vimentin (white) and Meis homeobox family (MEIS1/2/3, red). Images (C) and (D) are co-stained for proximal tubule marker lotus tetragonolobus lectin (green). All images show nuclei stained with DAPI (blue).

    Protocols and Documentation

    Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

    Document Type
    Product Name
    Catalog #
    Lot #
    Language
    Document Type
    Product Information Sheet
    Product Name
    STEMdiff™ Kidney Organoid Kit
    Catalog #
    05160
    Lot #
    All
    Language
    English
    Document Type
    Technical Manual
    Product Name
    STEMdiff™ Kidney Organoid Kit
    Catalog #
    05160
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 1
    Product Name
    STEMdiff™ Kidney Organoid Kit
    Catalog #
    05160
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 2
    Product Name
    STEMdiff™ Kidney Organoid Kit
    Catalog #
    05160
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 3
    Product Name
    STEMdiff™ Kidney Organoid Kit
    Catalog #
    05160
    Lot #
    All
    Language
    English

    Applications

    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.

    Research Area
    Workflow Stages
    Workflow Stages for Organoids
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    Resources and Publications

    Educational Materials (19)

    Organoids
    Brochure
    Organoids
    hPSC Differentiation: Tools for Pluripotent Stem Cell-Derived Research
    Brochure
    hPSC Differentiation: Tools for Pluripotent Stem Cell-Derived Research
    Products for Human Pluripotent Stem Cells
    Brochure
    Products for Human Pluripotent Stem Cells
    How to Prepare Single Cells from PSC-Derived Epithelial Cultures for Flow Cytometry Analysis
    Protocol
    How to Prepare Single Cells from PSC-Derived Epithelial Cultures for Flow Cytometry Analysis
    Performing Immunocytochemical Staining of PSC-Derived Kidney Organoids
    Protocol
    Performing Immunocytochemical Staining of PSC-Derived Kidney Organoids
    How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
    Protocol
    How to Process Epithelial Organoids and Organoid-Derived Epithelial Monolayers for RNA Isolation
    Derivation and Applications of Human Pluripotent Stem Cells
    Wallchart
    Derivation and Applications of Human Pluripotent Stem Cells
    Supporting Science Globally: A look into how scientists in Singapore are using organoids
    4:46
    Video
    Supporting Science Globally: A look into how scientists in Singapore are using organoids
    Collaborating to Accelerate COVID-19 Research
    3:36
    Video
    Collaborating to Accelerate COVID-19 Research
    HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
    1:10:09
    Webinar
    HUB & STEMCELL Organoids as Models of Infectious Disease Mini-Symposium
    Qualification of Ancillary/Raw Materials for Clinical Use
    54:39
    Webinar
    Qualification of Ancillary/Raw Materials for Clinical Use
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases, Nephrotoxicity, and Highly Relevant Cell-Based Assay
    59:52
    Webinar
    ISSCR Innovation Showcase: Applications of Organoid and Organotypic Cultures in Infectious Diseases,...
    Challenges in Translating iPSC Technology
    41:47
    Webinar
    Challenges in Translating iPSC Technology
    Re-Creating Disease with Kidney Organoids and CRISPR
    1:14:36
    Webinar
    Re-Creating Disease with Kidney Organoids and CRISPR
    Organoid Expert Panel
    52:35
    Webinar
    Organoid Expert Panel
    "COVID-19 Research Updates" Featuring Dr. Josef Penninger
    1:14:58
    Podcast
    "COVID-19 Research Updates" Featuring Dr. Josef Penninger
    High-Throughput-Compatible Differentiation of Human Pluripotent Stem Cell Lines to Kidney Organoids for Nephrotoxic Drug Screening
    Scientific Poster
    High-Throughput-Compatible Differentiation of Human Pluripotent Stem Cell Lines to Kidney Organoids ...
    Efficient, Reproducible and High-Throughput-Compatible Protocols for Differentiation of Human Pluripotent Stem Cell Lines Into Kidney Organoids
    Scientific Poster
    Efficient, Reproducible and High-Throughput-Compatible Protocols for Differentiation of Human Plurip...
    Dr. Bas Trietsch
    Interview
    Dr. Bas Trietsch
    PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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