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STEMdiff™ Endothelial Differentiation Kit

Efficient differentiation of human pluripotent stem cells to endothelial cells
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  • Efficient generation of hPSC-derived endothelial cells.

  • No cell enrichment or sorting step required.

  • Superior hPSC-derived endothelial expansion compared to FBS-contain media.

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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™ Endothelial Differentiation Kit (Catalog #08005) includes attachment substrate, animal component-free (ACF) endothelial induction medium, and endothelial expansion medium. It is optimized for the differentiation of human pluripotent stem cells (hPSCs) maintained in mTeSR™1 (Catalog #85850), mTeSR™ Plus (Catalog #05825), or TeSR™-E8™ (Catalog #05990) on Corning® Matrigel® to endothelial-like cells. The kit is designed to be used immediately after early mesoderm induction with STEMdiff™ Mesoderm Induction Medium (Catalog #05220), available for purchase separately.

    CollPlant is the manufacturer of the rhCollagen component of the cell attachment substrate.

    The Product may be used for research purposes only. Please contact STEMCELL for any clinical or commercial application of the Product.

    Data Figures

    Figure 1. Schematic Workflow of Endothelial Induction Using the STEMdiff™ Endothelial Kit

    In Phase 1, human embryonic stem (ES) or induced pluripotent stem (iPS) cells are cultured in a TeSR™ maintenance medium (mTeSR™ Plus, mTeSR™1, or TeSR™-E8™). On Day 1 (Phase 2) of the protocol, cells are ready for induction into early mesoderm progenitor cells by replacing TeSR™ medium with STEMdiff™ Mesodermal Induction Medium (MIM). By Day 3 (Phase 3), STEMdiff™ Mesoderm Induction Medium is replaced with STEMdiff™ Endothelial Induction Medium to derive endothelial cells. On Day 7, cells are passaged 5 - 6 times onto cultureware pre-coated with Animal Component-Free Cell Attachment Substrate in STEMdiff™ Endothelial Expansion Medium (Phase 4).

    Figure 2. A Representative Flow Cytometric Analysis of Endothelial Marker Expression in hPSC-Derived Endothelial Cells

    Human pluripotent stem cell (hPSC; H9 cell line)-derived endothelial cells were obtained at Day 7 using STEMdiff™ Endothelial Induction Medium. Greater than 85% of the cells were CD34+ and had high levels of CD31 and CD144 expression. With subsequent passages, the proportion of cells expressing endothelial markers (CD34+, CD31, and CD144) increased up to passage 5.

    Figure 3. STEMdiff™ Endothelial Differentiation Kit Generates Functional hPSC-Derived Endothelial Cells

    Endothelial cells generated from hPSCs (F016 cell line) using the STEMdiff™ Endothelial Differentiation Kit take up acetylated LDL when plated at 10,000 cells/cm2. Cells are able to form tubular networks in vitro in a tube formation assay when plated at 20,000 cells/well in a 96 well-plate for 24 hrs.

    Figure 4. Endothelial Cells Expand Faster in STEMdiff™ Endothelial Expansion Medium Compared to Serum-Containing Medium

    STEMdiff™ Endothelial Expansion Medium (A) sustains expansion rate in later passages and leads to (B) superior expansion of hPSC (C1 cell line)-derived endothelial cells when compared to serum-containing medium.

    Figure 5. hPSC-Derived Endothelial Cells Generated Using the STEMdiff™ Endothelial Differentiation Kit Express High Levels of ACE2

    (A) hPSC (C1 cell line)-derived endothelial cells were generated using the STEMdiff™ Endothelial Differentiation Kit and expanded in STEMdiff™ Endothelial Expansion Medium for 6 passages at 10,000 cells/cm2. (B) The cells were then analyzed for expression of angiotensin-converting enzyme 2 (ACE2). 85% of cells expressed high levels of ACE2.

    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™ Endothelial Differentiation Kit
    Catalog #
    08005
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 1
    Product Name
    STEMdiff™ Endothelial Differentiation Kit
    Catalog #
    08005
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 2
    Product Name
    STEMdiff™ Endothelial Differentiation Kit
    Catalog #
    08005
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 3
    Product Name
    STEMdiff™ Endothelial Differentiation Kit
    Catalog #
    08005
    Lot #
    All
    Language
    English
    Document Type
    Safety Data Sheet 4
    Product Name
    STEMdiff™ Endothelial Differentiation Kit
    Catalog #
    08005
    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
    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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