title: Mesenchymal differentiation propensity of a human embryonic stem cell line.
abstract: OBJECTIVE
  To characterize basal differentiation tendencies of a human embryonic stem (hES) cell line, KCL-002.
  
  
  METHODS
  In vitro specification and differentiation of hES cells were carried out using embryoid body (EB) cultures and tests of pluripotency and in vivo differentiation were performed by teratoma assays in SCID mice. Real-time PCR, immunohistochemistry, flow cytometry and histological analyses were used to identify expression of genes and proteins associated with the ectodermal, endodermal and mesodermal germ layers.
  
  
  RESULTS
  Undifferentiated KCL-002 cells expressed characteristic markers of pluripotent stem cells such as Nanog, Sox-2, Oct-4 and TRA 1-60. When differentiated in vitro as EB cultures, expression of pluripotency, endodermal and ectodermal markers decreased rapidly. In contrast, mesodermal and mesenchymal markers such as VEGFR-2, α-actin and vimentin increased during EB differentiation as shown by qPCR, immunostaining and flow cytometric analyses. Teratoma formation in SCID mice demonstrated the potential to form all germ layers in vivo with a greater proportion of the tumours containing mesenchymal derivatives.
  
  
  CONCLUSIONS
  The data presented suggest that the KCL-002 hES cell line is pluripotent and harbours a bias in basal differentiation tendencies towards mesodermal and mesenchymal lineage cells. Characterizing innate differentiation propensities of hES cell lines is important for understanding heterogeneity between different cell lines and for further studies aimed at deriving specific lineages from hES cells.
authors.affiliation: Stem Cell Laboratory, King's College London, London, UK.
authors.forename: S
authors.initials: S
authors.lastname: Pringle
authors.forename: C
authors.initials: C
authors.lastname: De Bari
authors.forename: F
authors.initials: F
authors.lastname: Dell'Accio
authors.forename: S
authors.initials: S
authors.lastname: Przyborski
authors.forename: M J
authors.initials: MJ
authors.lastname: Cooke
authors.forename: S L
authors.initials: SL
authors.lastname: Minger
authors.forename: A E
authors.initials: AE
authors.lastname: Grigoriadis
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D004475:Ectoderm / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004707:Endoderm / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D014183:Transplantation, Heterologous; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D000954:Antigens, Surface; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D011509:Proteoglycans; D055748:SOXB1 Transcription Factors; C107064:TRA-1-60 antigen, human; D040301:Vascular Endothelial Growth Factor Receptor-2