Assessment of Functional Competence of Endothelial Cells from Human Pluripotent Stem Cells in Zebrafish Embryos
Human pluripotent stem cells (hPSCs) are proving to be a valuable source of endothelial cells (ECs), pericytes, and vascular smooth muscle cells (vSMCs). Although an increasing number of phenotypic markers are becoming available to determine the phenotypes of these cells in vitro, the ability to integrate and form functional vessels in the host organism, typically mouse, remains critical for the assessment of EC functional competence. However, current mouse models require relatively large numbers of cells that might be difficult to derive simultaneously from multiple hPSCs lines. Therefore, there is an urgent need for new functional assays that are robust and can be performed with small numbers of cells. Here we describe a novel zebrafish xenograft model to test functionality of hPSC-derived ECs. The assay can be performed in 10 days and requires only ~100–400 human cells per embryo. Thus, the zebrafish xenograft model can be useful for the accurate and rapid assessment of functionality of hPSC-derived ECs in a lower vertebrate model that is widely viewed by regulatory authorities as a more acceptable alternative to adult mice.
Key wordsHuman pluripotent stem cells (hPSCs) Endothelial cells (ECs) Zebrafish xenograft
The authors would like to thank Francijna E. van den Hil for technical assistance.
We are grateful for financial support from LUMC (Gisela Thier Fellowship, V.V.O); the Netherlands Institute of Regenerative Medicine and the Netherlands Heart Foundation (2008B106) (C.LM.); Cancer Genomics Centre and LeDucq Foundation (P.D.).