Global transcriptomic analysis of murine embryonic stem cell-derived brachyury(+) (T) cells.
Brachyury(+) mesodermal cell population with purity over 79% was obtained from differentiating brachyury embryonic stem cells (ESC) generated with brachyury promoter driven enhanced green fluorescent protein and puromycin-N-acetyltransferase. A comprehensive transcriptomic analysis of brachyury(+) cells enriched with puromycin application from 6-day-old embryoid bodies (EBs), 6-day-old control EBs and undifferentiated ESCs led to identification of 1573 uniquely up-regulated and 1549 uniquely down-regulated transcripts in brachyury(+) cells. Furthermore, transcripts up-regulated in brachyury(+) cells have overrepresented the Gene Ontology annotations (cell differentiation, blood vessel morphogenesis, striated muscle development, placenta development and cell motility) and Kyoto Encyclopedia of Genes and Genomes pathway annotations (mitogen-activated protein kinase signaling and transforming growth factor beta signaling). Transcripts representing Larp2 and Ankrd34b are notably up-regulated in brachyury(+) cells. Knockdown of Larp2 resulted in a significantly down-regulation BMP-2 expression, and knockdown of Ankrd34b resulted in alteration of NF-H, PPARγ and PECAM1 expression. The elucidation of transcriptomic signatures of ESCs-derived brachyury(+) cells will contribute toward defining the genetic and cellular identities of presumptive mesodermal cells. Furthermore, there is a possible involvement of Larp2 in the regulation of the late mesodermal marker BMP-2. Ankrd34b might be a positive regulator of neurogenesis and a negative regulator of adipogenesis.
Doss
Michael Xavier
MX
Center of Physiology and Pathophysiology, Institute of Neurophysiology, and Center of Molecular Medicine, University of Cologne (CMMC), Robert-Koch Str. 39, 50931 Cologne, GermanyMax-Delbrueck-Center for Molecular Medicine - MDC, Robert-Rössle Str. 10, 13092 Berlin, GermanyInstitute of Computer Science, University of Tartu, Liivi 2, 50409 Tartu, Estonia and Quretec Ltd, Ulikooli 6a, Tartu, EstoniaDepartment of Vertebrate Genomics, Max-Planck-Institute for Molecular Genetics, Ihnestr.73, D-14195 Berlin, Germany.
Wagh
Vilas
V
Schulz
Herbert
H
Kull
Meelis
M
Kolde
Raivo
R
Pfannkuche
Kurt
K
Nolden
Tobias
T
Himmelbauer
Heinz
H
Vilo
Jaak
J
Hescheler
Jürgen
J
Sachinidis
Agapios
A
D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005326:Fetal Proteins / Q000378:metabolism*; D051379:Mice; D016900:Neurofilament Proteins / Q000378:metabolism; D047495:PPAR gamma / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D059467:Transcriptome; D000097585:SS-B Antigen; D000098908:Brachyury Protein
D000123:Acetyltransferases; D001324:Autoantigens; D055396:Bone Morphogenetic Protein 2; D005326:Fetal Proteins; D016900:Neurofilament Proteins; D047495:PPAR gamma; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D034741:RNA, Small Interfering; D012261:Ribonucleoproteins; D020825:T-Box Domain Proteins; D000097585:SS-B Antigen; D000098908:Brachyury Protein; C525719:Bmp2 protein, mouse; C051774:neurofilament protein H; C045336:puromycin N-acetyltransferase