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Generation of functional hemangioblasts from human embryonic stem cells

Nature Methods volume 4, pages 501–509 (2007)Cite this article

Abstract

Recent evidence suggests the existence of progenitor cells in adult tissues that are capable of differentiating into vascular structures as well as into all hematopoietic cell lineages. Here we describe an efficient and reproducible method for generating large numbers of these bipotential progenitors—known as hemangioblasts—from human embryonic stem (hES) cells using an in vitro differentiation system. Blast cells expressed gene signatures characteristic of hemangioblasts, and could be expanded, cryopreserved and differentiated into multiple hematopoietic lineages as well as into endothelial cells. When we injected these cells into rats with diabetes or into mice with ischemia-reperfusion injury of the retina, they localized to the site of injury in the damaged vasculature and appeared to participate in repair. Injection of the cells also reduced the mortality rate after myocardial infarction and restored blood flow in hind limb ischemia in mouse models. Our data suggest that hES-derived blast cells (hES-BCs) could be important in vascular repair.

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Figure 1: Characterization of hES-BCs.
Figure 2: Clonogenicity of blast colonies.
Figure 3: Repair of ischemic retinal vasculature in a mouse after injection of hES-BCs.
Figure 4: Incorporation of hES-BCs into the retinal vasculature of diabetic rats.
Figure 5: Endothelial differentiation in infarcted heart and in ischemic hind limb muscle after injection of hES-BCs.

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Acknowledgements

We thank Y. Ivanova, S. Becker, T.Li, C. Luo, I. Klimanskaya (Advanced Cell Technology) and N. Sengupta (University of Florida) for their excellent technical assistance; The Juvenile Diabetes Research Foundation International; NIH grants EY012601; EY007739; EY14818 (to M.B.G.).

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Authors and Affiliations

  1. Advanced Cell Technology, Worcester, 01605, Massachusetts, USA

    Shi-Jiang Lu, Qiang Feng & Robert Lanza

  2. Program in Stem Cell Biology and Department of Pharmacology and Therapeutics, University of Florida, Gainesville, 32610, Florida, USA

    Sergio Caballero & Maria B Grant

  3. Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    Yu Chen & Malcolm A S Moore

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  1. Shi-Jiang Lu
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  7. Robert Lanza
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Corresponding author

Correspondence to Robert Lanza.

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Competing interests

S.J.L., Q.F. and R.L. are employees of Advanced Cell Technology (ACT), a stem cell company in the field of regenerative medicine; M.A.S.M. is on ACT's Scientific Advisory Board.

Supplementary information

Supplementary Fig. 1

Characterization of hES-BC cells derived from hES cells (PDF 166 kb)

Supplementary Fig. 2

Hematopoietic CFUs derived from hES-BC cells (PDF 126 kb)

Supplementary Fig. 3

FACS analysis of hematopoietic cells of pooled CFUs derived from hES-BC cells (PDF 158 kb)

Supplementary Fig. 4

Endothelial cells derived from hES-BC cells (PDF 119 kb)

Supplementary Fig. 5

PCR analysis of GFP sequences in GFP+ and GFP− hES-BCs derived from plating mixtures of WA01-GFP+ and MA01-GFP− cells (PDF 121 kb)

Supplementary Table 1

Efficiency of hES BC cells derived from different hES cell lines (PDF 60 kb)

Supplementary Methods (PDF 135 kb)

Supplementary Protocol

Protocol for culturing, differentiating and expanding hES-BC cells. (PDF 94 kb)

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Lu, SJ., Feng, Q., Caballero, S. et al. Generation of functional hemangioblasts from human embryonic stem cells. Nat Methods 4, 501–509 (2007). https://doi.org/10.1038/nmeth1041

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