Human iPSC-Derived Endothelial Colony Forming Cells

Endothelial Colony Forming Cells

Endothelial colony forming cells (ECFCs) are rare, highly proliferative endothelial cells that are found in bone marrow and the bloodstream. ECFCs are highly angiogenic and can migrate to sites of blood vessel injury and contribute to repair of the damaged vessels. In partnership with Professor Mervin Yoder's laboratory at Indiana University, Axol has developed iPSC-Derived Endothelial Colony Forming Cells that resemble primary ECFCs in their cell surface marker expression and functionality.
Axol iPSC-Derived Endothelial Colony Forming Cells express the endothelial markers CD31, CD144, KDR and NRP1. The endothelial cells align in response to unidirectional shear flow and are responsive to pro-angiogenic factors secreted by fibroblasts. The ECFCs are stable and, unlike other iPSC-derived endothelial cell models, they do not transition to non-endothelial cell types during long-term culture. The endothelial colony forming cells can be passaged up to 10 times, permitting high throughput experiments that require large quantities of cells from the same donor. Axol iPSC-Derived  ECFCs have comparable cell marker expression to primary HUVECs and have the advantage of greater consistency and the possibility of performing co-culture experiments with other cell types derived from the same iPSCs such as neural stem cells.


Human iPSC-derived endothelial colony forming cells 

Endothelial colony forming cells from healthy male and female donor iPSCs. The cells are cryopreserved at passage 4 and are provided at 1 million viable cells per vial.  Axol iPSC-Derived Endothelial Colony Forming Cells can be stably expanded and repeatedly passaged in Axol Endothelial Colony Forming  Cell Culture Medium.

Cat. No. Product Name Starting Material Quantity Price
ax2015 Human iPSC-Derived Endothelial Colony Forming Cells (Male) Cord Blood CD34+ Cells 1 million cells $960 (Pre-order now)
ax2019 Human iPSC-Derived Endothelial Colony Forming Cells (Female) Fibroblasts (64 Yr Donor) 1 million cells $960 (Pre-order now)

Characterization

Endothelial cell marker expression

Expression of the endothelial cell-specific markers: von Willebrand factor, endothelial NOS and CD31 in Axol Human iPSC-Derived Endothelial Colony Forming Cells (ECFCs). Hoechst 33342 was used to visualize cell nuclei. The images were kindly provided by Dr Richard Siow (King's College London).

Expression of endothelial and lymph marker proteins in Axol iPSC-Derived Endothelial Colony Forming Cells as determined by flow cytometry. The cells demonstrate a clear endothelial cell phenotype and the expression of LYVE1 suggests the potential for differentiation to lymphatic endothelium. The dashed lines indicate negative control fluorescence levels and the solid lines represent the fluorescence of stained iPSC-Derived Endothelial Colony Forming Cells . The data were kindly provided by Dr Wolfgang Holnthoner (Ludwig-Boltzmann-Institute for Experimental and Clinical Traumatology).


Co-culture

Co-culture of iPSC-Derived Endothelial Colony Forming Cells with lung fibroblasts to assess angiogenic potential

Axol iPSC-Derived Endothelial Colony Forming Cells  (iPSC-ECFC) were seeded into a co-culture vessel together with human lung fibroblasts. By day 4, the endothelial cells grow towards the lung fibroblasts in response to secreted angiogenic factors. The process can be used to model the formation of perfusable blood vessels during angiogenesis. The images were kindly provided by Dr Jungho Ahn (Seoul National University).

Axol iPSC-Derived Endothelial Colony Forming Cells extend outgrowths towards human lung fibroblasts when co-cultured. Primary human umbilical vein endothelial cells (HUVECs) were included as a positive control. The outgrowths were fixed and stained for CD31 (red), F-actin (green) and DAPI (blue). Scale bar = 150 μm. The images were kindly provided by Dr Jungho Ahn (Seoul National University).

Co-culture of iPSC-Derived Endothelial Colony Forming Cells with adipose tissue-derived stem cells to assess formation of vascular networks

Axol iPSC-Derived Endothelial Colony Forming Cells were co-cultured with adipose tissue-derived stem cells in a fibrin clot (fibrinogen + thrombin). After 7 days, the cells have started to form a vascular network. The fibrin clots were fixed and stained for CD31 (green). The images were kindly provided by Dr Wolfgang Holnthoner (Ludwig-Boltzmann-Institute for Experimental and Clinical Traumatology).


Endothelial cell alignment

Response of iPSC-Derived Endothelial Colony Forming Cells to shear flow

ECFC-shear-force-alignment.jpg

Axol iPSC-Derived Endothelial Colony Forming Cells were cultured in microfluidic slides under oscillatory or unidirectional shear flow. The endothelial cells responded to unidirectional shear flow by aligning. In static and oscillatory shear conditions, the endothelial cells did not align. The images were kindly provided by Dr Richard Siow (King's College London).


Citations