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


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 48 hours.


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.
Hill
Katherine L
KL
Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, Minn. 55455, USA.
Obrtlikova
Petra
P
Alvarez
Diego F
DF
King
Judy A
JA
Keirstead
Susan A
SA
Allred
Jeremy R
JR
Kaufman
Dan S
DS
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D001808:Blood Vessels / Q000166:cytology*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D019070:Cell Lineage; D002478:Cells, Cultured; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D005434:Flow Cytometry; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism
D018952:Antigens, CD34; D008840:Microfilament Proteins; D009124:Muscle Proteins; D051153:Wnt Proteins; C051615:transgelin