Cell fusion-independent differentiation of neural stem cells to the endothelial lineage

Abstract

Somatic stem cells have been claimed to possess an unexpectedly broad differentiation potential (referred to here as plasticity) that could be induced by exposing stem cells to the extracellular developmental signals of other lineages in mixed-cell cultures1,2,3,4,5,6. Recently, this and other experimental evidence supporting the existence of stem-cell plasticity have been refuted because stem cells have been shown to adopt the functional features of other lineages by means of cell-fusion-mediated acquisition of lineage-specific determinants (chromosomal DNA) rather than by signal-mediated differentiation1,2,5,7,8. In this study we co-cultured mouse neural stem cells (NSCs), which are committed to become neurons and glial cells9,10, with human endothelial cells, which form the lining of blood vessels11. We show that in the presence of endothelial cells six per cent of the NSC population converted to cells that did not express neuronal or glial markers, but instead showed the stable expression of multiple endothelial markers and the capacity to form capillary networks. This was surprising because NSCs and endothelial cells are believed to develop from the ectoderm and mesoderm, respectively. Experiments in which endothelial cells were killed by fixation before co-culture with live NSCs (to prevent cell fusion) and karyotyping analyses, revealed that NSCs had differentiated into endothelial-like cells independently of cell fusion. We conclude that stem-cell plasticity is a true characteristic of NSCs and that the conversion of NSCs to unanticipated cell types can be accomplished without cell fusion.

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Figure 1: NSCs can be induced to express the endothelial marker, CD146.
Figure 2: Endothelial markers are expressed by NSC-derived endothelial cell candidates clonally purified from human endothelial cell co-culture.
Figure 3: NSC-derived endothelial cell candidates show endothelial Weibel–Palade bodies and form capillary networks in vitro.
Figure 4: Cell fusion is unlikely to contribute to the generation of NSC-derived endothelial-like cells.

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Acknowledgements

We thank M. G. Farquhar and R. V. Stan for critically evaluating Weibel–Palade bodies; D. Chambers for conducting FACS analysis; J. Simon for help in constructing figures; L. Moore, B. Miller, A. Hyunh and S. Thuret for technical assistance; J. D. Esko's laboratory for advice; V. A. Sciorra and M. L. Gage for critical reading of the manuscript. This work was supported by the National Institutes of Health (National Institute on Aging) and the Lookout Fund. A.E.W. is a Damon Runyon fellow supported by The Damon Runyon Cancer Research Foundation. K.N. was supported by a JSPS Postdoctoral Fellowship for Research Abroad. F.H.G. is supported by the Vi and John Adler Chair.

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Correspondence to Fred H. Gage.

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While I do not believe there is a real financial interest, there could be a perceived competing interest with three companies that I am on the SAB for: CEREGENE inc., Stem Cells inc. and B.C. inc.

Supplementary information

Supplementary Figure 1

Documents the expression of multiple endothelial markers by NSC-derived ECs using immunofluorescence and RT-PCR. (JPG 64 kb)

Supplementary Movie

NSC-derived EC marker-expressing cells form capillary networks in vitro. NSCs, human ECs and NSC-derived EC candidates (clone 1 is shown) were introduced onto Matrigel coated plates at the 0 h time point and monitored for the capacity to form vascular cords. Scale bar=100 µm. (MOV 2657 kb)

Supplementary Figure and Movie Legend (DOC 20 kb)

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Wurmser, A., Nakashima, K., Summers, R. et al. Cell fusion-independent differentiation of neural stem cells to the endothelial lineage. Nature 430, 350–356 (2004). https://doi.org/10.1038/nature02604

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