iCell Endothelial Cells, 11713

Kit Size

Catalog #: R1112

Cells Only

Catalog #: C1114
Catalog #: C1114

Endothelial Cells differentiated from human iPS cells, frozen

From
$1,025.00
Catalog # GETC11713

Product Overview

Endothelial cells are highly dynamic cells that form the inner lining of blood vessels. In addition to their structural role, endothelial cells regulate the passage of substances between the bloodstream and surrounding tissue. They also secrete mediators that influence vascular hemodynamics, inflammation, cell trafficking, and the remodeling and formation of new blood vessels. Endothelial cell dysfunction is associated with a variety of diseases including atherosclerosis, coronary artery disease, hypertension, and inflammatory disorders.

Derived from human iPS cells, FCDI’s iCell® Endothelial Cells provide a reliable, reproducible, and physiologically relevant source of cells for vascular disease modeling, drug screening, and vascular tissue engineering applications. Benefits include:

  • >90% CD31+/CD105+ populations across lots and multiple passages
  • VEGF-mediated proliferation, sprouting, migration, and invasion
  • TNFα-mediated upregulation of adhesion molecules
  • Thrombin-sensitive barrier function
  • Acetylated LDL uptake
  • Shear stress-induced alignment
  • 2D and 3D capillary-like network formation

Broad Function and Application

Assayed as a pure culture or in co-culture with other differentiated cell types to recapitulate native tissue architecture, iCell Endothelial Cells enable a variety of novel applications:

  • Vascular tissue engineering:
    Decellularized organs Amenable to decellularized organ (e.g. kidney) scaffold approaches (Caralt et al., 2015)
  • Vascular tissue engineering:
    Natural matrices and bioengineering scaffolds Compatible with natural matrix approaches as well as microfluidic device and bioreactor system cultures (Bischel et al. 2014; Belair et al., 2014)
  • Angiogenesis and wound healing
    Providing relevant biology for the study of blood vessel formation and remodeling relevant to cancer, chronic inflammatory, and degenerative diseases
  • Infectious disease studies
    Serving as a novel human in vitro model for the study of bacterial and viral infections

Components:

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Performance Data

Formation of 3D Cardiac Tri-Culture Microtissues

Three-dimensional multi-cellular systems containing iCell Cardiomyocytes or iCell Cardiomyocytes2, iCell Endothelial Cells and primary cardiac fibroblasts have the potential for greater physiological relevance, predictive power, and mechanistic insight than cardiomyocytes alone. For information on 3D systems, see the Application Protocol: Culturing and Assaying Calcium Transients of 3D Cardiac Tri-Culture Microtissues

Figure 1: Structure of 3D Cardiac Tri-culture Microtissues Over Time.
Microtissues were formed containing 5,000 or 10,000 total cells in S-bio 96-well plates. Compact, contracting microtissues were obtained by Day 4. (A) Panel of phase contrast 10X images of 5,000 cell tri-culture spheroids over time using the Incucyte S3. (B) Quantification of 5,000 cell tri-culture microtissue diameter. Each dot represents a microtissue. Mean and SEM are indicated. (C) Comparison of Day 14 microtissue diameter of 5,000 cell tri-culture microtissues with 10,000 cell tri-culture microtissues. Each dot represents a microtissue. Mean and SEM are indicated. (D) H&E staining of Day 14, 3D cardiac tri-culture microtissue formed with 10,000 total cells. Staining shows the absence of a necrotic core.

Figure 2: 3D Cardiac Tri-culture Microtissues Response to Beta Adrenergic Agonist Isoproterenol
(A) Chronotropic response: Control iCell Cardiomyocytes, 11713 only microtissues (CM) and tri-culture microtissues exhibit an increase in beat rate with increasing concentrations of isoproterenol. (B) Inotropic response: Control iCell Cardiomyocytes, 11713 only microtissues (CM) do not increase amplitude with increasing concentrations of isoproterenol. Tri-culture microtissues demonstrate a twofold increase in beat amplitude with increasing concentrations of isoproterenol.

Enhanced Inotropic Response when Plated in Tri-Culture

3D cardiac tri-culture microtissues containing iCell Cardiomyocytes or iCell Cardiomyocytes2, iCell Endothelial Cells and cardiac fibroblasts demonstrated a positive response to the inotropic compound isoproterenol, which is characteristic of mature cardiomyocytes. For information on 3D triculture systems, see the Application Protocol: Culturing and Assaying Calcium Transients of 3D Cardiac Tri-Culture Microtissues

Compatible with High-Sensitivity Detection of Calcium Transients in Tri-Culture

Three-dimensional multi-cellular systems containing iCell Cardiomyocytes or iCell Cardiomyocytes2, iCell Endothelial Cells and cardiac fibroblasts demonstrate enhanced amplitude in calcium transient assays. For information on 3D systems, see the Application Protocol: Culturing and Assaying Calcium Transients of 3D Cardiac Tri-Culture Microtissues

Figure 3: Baseline Calcium Transients in Cardiomyocyte and Tri-culture Microtissues at Day 14
Calcium transients were measured using EarlyTox calcium dye. (A) Representative calcium traces for 5,000 and 10,000 total cell 3D Tri-culture cardiac microtissues. (B) Beat rate is not different between 5,000 or 10,000 cell triculture microtissues and control cardiomyocyte only microtissues. (C) Representative calcium traces for 5,000 cell tri-culture and 5,000 cell cardiomyocytes only microtissues (CM only). (C) Amplitude is significantly higher in triculture microtissue compared to cardiomyocyte only microtissues (CM only) at 5,000 cells, but similar at 10,000 cells

Figure 4: iCell Endothelial Cells Are a Highly Pure Population

iCell Endothelial Cells Purity Demonstrated by Flow Cytometry Analysis

The cells have >90% expression of CD105 (endoglin), CD31 (PECAM-1) and CD144 (VE-cad) as determined by flow cytometry analysis.

iCell Endothelial Cell Markers and Functions Assessed by Immunostaining

(A) When immunostained for von Willebrand Factor (vWF), the cells showed characteristic Weibel-Palade body staining. (B) In a thick layer of Matrigel, the cells exhibited the capacity to form tubes. (C) To assess the barrier function, the cells were immunostained for the tight junction protein ZO-1.

Figure 5: iCell Endothelial Cells Display Expected Markers and Functions

Figure 6: Barrier Disruption Effect of Thrombin on iCell Endothelial Cell

Barrier Function Assay Using the xCELLigence RTCA Platform

iCell Endothelial Cells exhibit barrier function activity that can be disrupted by treatment with thrombin and reliably assessed using impedance-based platforms. iCell Endothelial Cells provide an in vitro test system that recapitulates native human endothelium properties and functions while the xCELLigence RTCA Cardio System provides a label-free technology for non-invasive monitoring of cell behavior and viability. The methods and results presented here highlight how to gather relevant data on human endothelial viability and barrier function. Together, iCell Endothelial Cells and impedance-based technologies offer a valuable cell model system for understanding the endothelial barrier characteristics, mechanisms of endothelial barrier dysfunction, and dynamic modulation of the endothelium permeability, enabling a wide range of applications in academic and pharmaceutical research. iCell Endothelial Cells were cultured for 48 hours on the E-Plate in Complete iCell Endothelial Cells Maintenance Medium before the addition of thrombin. (A) The representative Cell Index curves showed the dose-dependent disruption effect of thrombin on the barrier function and the recovery phase. (B) The percentage of Cell Index compared to pre-treatment baseline levels was calculated at the time of thrombin addition, at 9 hours post-treatment, or at 21 hours post-treatment (mean ± SD, n = 2 wells).

Product Highlights

Applications

iCell Endothelial Cells are amenable to a variety of assays including:

  • Angiogenesis/Vasculogenesis
  • Cell permeability
  • Cell adhesion
  • Cell proliferation
  • Cell invasion
  • Impedance/Barrier function
  • Cell migration
  • Tube formation

Advantages:

  • Human cells: iCell Endothelial Cells are differentiated from human iPS cells and exhibit characteristics and functions of endothelial cells.
  • .
  • Homogenous and reproducible: iCell Endothelial Cells are highly pure, providing biologically relevant and reproducible results.
  • Acute and long-term testing: iCell Endothelial Cells actively proliferate, remaining viable and pure in culture for weeks, enabling assessment of both acute and sub-chronic responses.
  • Easy to implement: iCell Endothelial Cells are shipped cryopreserved with cell culture medium supplement specifically formulated for optimal cell performance. Simply thaw and use.

Publications