Identification of cardiac stem cells with FLK1, CD31, and VE-cadherin expression during embryonic stem cell differentiation.
We evaluated the expression of the FLK1, one of the lateral mesoderm early markers where cardiogenesis occurs, to characterize and isolate cardiac stem/progenitor cells from ES cells. Dissociated cells from embryoid bodies (EBs) on day 3, 4, or 5 were collected into two subpopulations with or without FLK1 expression and coculture on OP9 stromal cells was continued to examine whether contracting colonies came out or not. FLK1+ cells from EBs at days 3 and 4 formed spontaneous contracting colonies more efficiently than FLK1- cells on the same days, but not at day 5. Most contracting cardiac colonies derived from FLK1+cells mainly on day 4 were detected on endothelial cells along with hematopoietic cells. Further characterization of cells with these capabilities into three lineages revealed the FLK1+ CD31-VE-cadherin-phenotype. Our findings indicate that FLK1+cells, especially FLK1+ CD31-VE-cadherin-cells, could act as cardiohemangioblasts to form cardiac cells as well as endothelial cells and hematopoietic cells.
Iida
Midori
M
Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto city, Kyoto, Japan.
Heike
Toshio
T
Yoshimoto
Momoko
M
Baba
Shiro
S
Doi
Hiraku
H
Nakahata
Tatsutoshi
T
D015703:Antigens, CD; D015820:Cadherins / Q000032:analysis*; D002454:Cell Differentiation; D002460:Cell Line; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000166:cytology*; D004625:Embryo, Nonmammalian; D042783:Endothelial Cells / Q000737:chemistry; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000737:chemistry; D007150:Immunohistochemistry; D009206:Myocardium / Q000166:cytology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000737:chemistry* / Q000166:cytology; D013997:Time Factors; D014162:Transfection; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis* / Q000235:genetics; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D049452:Green Fluorescent Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D000099152:Cadherin 5; C485184:enhanced green fluorescent protein