STEMdiff™ Endothelial Differentiation Kit

Efficient differentiation of human pluripotent stem cells to endothelial cells
STEMdiff™ Endothelial Differentiation Kit

Efficient differentiation of human pluripotent stem cells to endothelial cells

1 Kit
Catalog # 08005
Required Products
  1. mTeSR™1
    mTeSR™1

    cGMP, feeder-free maintenance medium for human ES and iPS cells

  2. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  3. STEMdiff™ Mesoderm Induction Medium|05220
    STEMdiff™ Mesoderm Induction Medium

    Defined, xeno-free induction medium for early mesodermal differentiation

  4. Heparin Solution
    Heparin Solution

    Cell culture supplement

Overview

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.
Advantages
⦁ Efficient generation of hPSC-derived endothelial cells.

⦁ No cell enrichment or sorting step required.

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

Components
  • STEMdiff™ Endothelial Induction Medium Kit (Catalog #08005)
    • STEMdiff™ Endothelial Induction Medium, 100 mL
    • STEMdiff™ Endothelial Expansion Basal Medium, 120 mL
    • STEMdiff™ Endothelial Expansion 5X Supplement, 30 mL
    • Animal Component-Free Cell Attachment Substrate, 1 mL
Contains
Cell Culture Media and Supplements
Subtype
Specialized Media
Cell Type
Endothelial Cells
Species
Human
Application
Differentiation
Brand
STEMdiff
Area of Interest
Disease Modeling, Endothelial Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

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

Educational Materials (6)

Brochure
Products for Human Pluripotent Stem Cells
Wallchart
Derivation and Applications of Human Pluripotent Stem Cells
Webinar
Therapeutic Application of Endothelial Colony-Forming Cells for Retinal Diseases
29:38
Therapeutic Application of Endothelial Colony-Forming Cells for Retinal Diseases
Mini Review
Endothelial Cells, Angiogenesis, and Vasculogenesis
Mini Review
Endothelial Progenitor Cells and Endothelial Cells
Scientific Poster
STEMdiff™ Endothelial: A New Culture Workflow for Efficient Derivation and Expansion of Endothelial Cells From Human Pluripotent Stem Cells

Product 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.

Data and Publications

Data

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.

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