In vitro development of a hemangioblast from a human embryonic stem cell, SNUhES#3.
OBJECTIVE
Recent reports demonstrated that a hemangioblast population emerged during hematopoietic development in both mouse and human embryonic stem cell (hESC) differentiation cultures.


METHODS
In this study, a new uncharacterized hESC line, SNUhES#3, was studied for its capacity to proliferate with STO cells and differentiate into hemangioblasts in co-culture with OP9 cells.


RESULTS
We were able to obtain CD34(+)CD45(-) cells from SNUhES#3 cells after 12 days of in vitro culture, and this cell population could be maximized to 12.6% of the total. These cells, derived from SNUhES#3, showed the morphology of hematopoietic precursor cells and endothelial lineage cells with high efficiency. Reverse transcription polymerase chain reaction (RT-PCR) analysis showed that the hematopoietic markers CD34, GATA2, and LMO2 were co-expressed with the endothelial marker CD31 from day 8, whereas ES cell marker OCT4 no longer existed at an early stage. Moreover, we found that the efficacy of colony forming by SNUhES#3 cells is better than that of H9 cells.


CONCLUSIONS
These findings provide evidence that SNUhES#3 cells can be used as an established human ESC line, and co-culture with OP9 can induce SNUhES#3 cells to differentiate into hemangioblasts, the common precursors of the hematopoietic and endothelial lineages.
Choi
Jae-Hyeog
JH
IRC, Department of Biological Sciences, University of Ulsan, Ulsan, Republic of Korea.
Ryu
Young-Shin
YS
Kim
Koo-Hee
KH
Lee
Yoo-Ra
YR
Cha
Ki-Won
KW
Han
In-Seob
IS
Kwon
Byoung S
BS
D018952:Antigens, CD34 / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology*; D005434:Flow Cytometry; D015870:Gene Expression / Q000235:genetics / Q000502:physiology; D005819:Genetic Markers; D055018:Hemangioblasts / Q000502:physiology*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000235:genetics; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells
D018952:Antigens, CD34; D005819:Genetic Markers; D017493:Leukocyte Common Antigens