Different differentiation kinetics of vascular progenitor cells in primate and mouse embryonic stem cells.
BACKGROUND
We demonstrated that vascular endothelial growth factor receptor 2 (VEGF-R2)-positive cells derived from mouse embryonic stem (ES) cells can differentiate into both endothelial cells and mural cells to suffice as vascular progenitor cells (VPCs). Here we examined whether VPCs occur in primate ES cells and investigated the differences in VPC differentiation kinetics between primate and mouse ES cells.


RESULTS
In contrast to mouse ES cells, undifferentiated monkey ES cells expressed VEGF-R2. By culturing these undifferentiated ES cells for 4 days on OP9 feeder layer, VEGF-R2 expression disappeared, and then reappeared after 8 days of differentiation. We then isolated these VEGF-R2-positive and vascular endothelial cadherin (VEcadherin)-negative cells by flow cytometry sorting. Additional 5-day reculture of these VEGF-R2+ VEcadherin- cells on OP9 feeder layer resulted in the appearance of platelet endothelial cell adhesion molecule-1 (PECAM1)-positive, VEcadherin-positive, endothelial nitric oxide synthase (eNOS)-positive endothelial cells. On a collagen IV-coated dish in the presence of serum, these cells differentiated into smooth muscle actin (SMA)-positive and calponin-positive mural cells (pericytes or vascular smooth muscle cells). Addition of 50 ng/mL VEGF to the culture on a collagen IV-coated dish resulted in the appearance of PECAM1+ cells surrounded by SMA+ cells. In addition, these differentiated VEGF-R2+ cells can form tube-like structures in a 3-dimensional culture.


CONCLUSIONS
Our findings indicate that differentiation kinetics of VPCs derived from primate and mouse ES cells were different. Differentiated VEGF-R2+ VEcadherin- cells can act as VPCs in primates. To seek the clinical potential of VPCs for vascular regeneration, investigations of primate ES cells are indispensable.
Sone
Masakatsu
M
Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507 Japan.
Itoh
Hiroshi
H
Yamashita
Jun
J
Yurugi-Kobayashi
Takami
T
Suzuki
Yutaka
Y
Kondo
Yasushi
Y
Nonoguchi
Akane
A
Sawada
Naoki
N
Yamahara
Kenichi
K
Miyashita
Kazutoshi
K
Park
Kwijun
K
Shibuya
Masabumi
M
Nito
Shinji
S
Nishikawa
Shin-Ichi
S
Nakao
Kazuwa
K
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004622:Embryo, Mammalian / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology*; D007700:Kinetics; D008252:Macaca fascicularis; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D013045:Species Specificity; D013234:Stem Cells / Q000737:chemistry / Q000502:physiology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis
D040301:Vascular Endothelial Growth Factor Receptor-2