Human embryonic stem cell−derived vascular progenitor cells capable of endothelial and smooth muscle cell function

Hill, Katherine L.; Obrtlikova, Petra; Alvarez, Diego F.; King, Judy A.; Keirstead, Susan A.; Allred, Jeremy R.; Kaufman, Dan S.

doi:10.1016/j.exphem.2010.01.001

« PreviousExperimental Hematology
Volume 38, Issue 3 , Pages 246-257.e1, March 2010

Human embryonic stem cell−derived vascular progenitor cells capable of endothelial and smooth muscle cell function

Katherine L. Hill
- Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn., USA
- Drs. Hill and Obrtlikova contributed equally to this work as co−first authors.
Petra Obrtlikova
- Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn., USA
- Clinical Department of Hematology of First Faculty of Medicine and General Teaching Hospital, Charles University, Prague, Czech Republic
- Drs. Hill and Obrtlikova contributed equally to this work as co−first authors.
Diego F. Alvarez
- Department of Internal Medicine, Pharmacology, Center for Lung Biology, University of South Alabama College of Medicine, Mobile, Ala., USA
Judy A. King
- Department of Internal Medicine, Pharmacology, Center for Lung Biology, University of South Alabama College of Medicine, Mobile, Ala., USA
Susan A. Keirstead
- Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn., USA
Jeremy R. Allred
- Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn., USA
Dan S. Kaufman
- Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn., USA
- Offprint requests to: Dan S. Kaufman, M.D., Ph.D., Stem Cell Institute and Department of Medicine, University of Minnesota, Masonic Cancer Center, 525 East River Parkway, Minneapolis, MN 55455

Received 13 September 2009; received in revised form 31 December 2009; accepted 4 January 2010. published online 11 January 2010.

Objective

Previous studies have demonstrated development of endothelial cells (ECs) and smooth muscle cells (SMCs) as separate cell lineages derived from human embryonic stem cells (hESCs). We demonstrate CD34⁺ cells isolated from differentiated hESCs function as vascular progenitor cells capable of producing both ECs and SMCs. These studies better define the developmental origin and reveal the relationship between these two cell types, as well as provide a more complete biological characterization.

Materials and Methods

hESCs are cocultured on M2-10B4 stromal cells or Wnt1-expressing M2-10B4 for 13 to 15 days to generate a CD34⁺ cell population. These cells are isolated using a magnetic antibody separation kit and cultured on fibronectin-coated dishes in EC medium. To induce SMC differentiation, culture medium is changed and a morphological and phenotypic change occurs within 24 to 48hours.

Results

CD34⁺ vascular progenitor cells give rise to ECs and SMCs. The two populations express respective cell-specific transcripts and proteins, exhibit intracellular calcium in response to various agonists, and form robust tube-like structures when cocultured in Matrigel. Human umbilical vein endothelial cells cultured under SMC conditions do not exhibit a change in phenotype or genotype. Wnt1-overexpressing stromal cells produced an increased number of progenitor cells.

Conclusions

The ability to generate large numbers of ECs and SMCs from a single vascular progenitor cell population is promising for therapeutic use to treat a variety of diseased and ischemic conditions. The stepwise differentiation outlined here is an efficient, reproducible method with potential for large-scale cultures suitable for clinical applications.