Identification of differentially expressed genes in human embryonic stem cell-derived endothelial cells using suppression subtractive hybridization.
Human embryonic stem cells (hESCs) are pluripotent, self-renewing cells derived from the inner cell mass of human blastocysts. During normal development, hESCs differentiate into 3 germ layers. Cellular lineages differentiated from hESCs express a set of genes that are exclusive to these specialized cells. Therefore, we hypothesized that endothelial cells derived from hESCs would express genes specific to endothelial cells. We previously isolated endothelial cells from human embryonic stem cells (hESC-ECs) using fluorescence-activated cell sorter (FACS). The aim of the current study was to identify genes associated with hESC-derived endothelial-like cells. Using suppression subtractive hybridization (SSH), we identified a set of genes specific to cells differentiated from hESC-ECs. We obtained 113 clones of expressed sequences that were more abundant in hESC-ECs compared with hESCs. Based on the NCBI GenBank database, 56 of these clones were known genes, 13 clones corresponded to nucleotides, 2 clones showed homology with chromosome sequences, and 42 clones showed no significant homology with any nucleotide sequences. These identified genes are related to cytoskeleton and cell adhesion, development, heat shock protein, metabolism, signal transduction, and transcription/nuclear-specific proteins. We anticipate that further study of these genes will provide crucial insights into their specific roles in the development of endothelial cells from hESCs.
Ahn
Jin Seop
JS
Department of Biomedical Science, College of Life Science, CHA University, Pochon-si, Gyeonggi-do, Korea.
Moon
Sung-Hwan
SH
Kim
Jumi
J
Chung
Hyung-Min
HM
Kim
Jin Kyeoung
JK
D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D048430:Cell Shape; D018076:DNA, Complementary / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D018919:Neovascularization, Physiologic; D009693:Nucleic Acid Hybridization / Q000379:methods; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics; D042787:Receptor, TIE-2 / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015854:Up-Regulation / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics; D014841:von Willebrand Factor / Q000378:metabolism
D018076:DNA, Complementary; D008077:Lipoproteins, LDL; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C032629:acetyl-LDL; D014841:von Willebrand Factor; D042787:Receptor, TIE-2; D040301:Vascular Endothelial Growth Factor Receptor-2