Expression of transcription factor Oct-4 and other embryonic genes in CD133 positive cells from human umbilical cord blood.
A significant number of hematopoietic stem/progenitor cells (HSPC) in human umbilical cord blood could serve as a reservoir for the placental vasculature, yet, their morphological and functional features are not completely understood. Here, we describe the characterization of purified CD133(+) progenitor cells from umbilical cord blood, a subset of CD34(+) hematopoietic progenitors that were grown in proliferation medium containing Flt3-ligand, thrombopoietin and stem cell factor. Following isolation and enrichment of the CD133(+) cells by immunomagnetic cell sorting, they remained non-adherent for up to 40 days in culture and expressed different pluripotency markers including Sox-1, Sox-2, FGF-4, Rex-1 and Oct-4.Oct-4 expression was confirmed by laser-assisted single cell picking with subsequent quantitative real-time RT-PCR. The expression of Oct-4 indicates a pluripotent phenotype of CD133(+) cells and appears to be of functional relevance: After three weeks in endothelial differentiation medium, suspended cells became adherent, developed an endothelial cell-like morphology, bound fluoresceine isothiocyanate-labeled Ulex europaeus agglutinin-1, took up acetylated Di-LDL, and expressed other endothelial markers such as PECAM-1 or VEGFR-2. Concomitantly, Oct-4 expression was significantly reduced. Moreover, following treatment with retinoic acid, CD133(+) cells exhibited neural morphology associated with the expression of beta-III-tubulin. CD133(+) cells were found to express the luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor, detected by RT-PCR and immunocytochemistry. The recombinant human chorionic gonadotropin induced proliferation of the CD133(+) cells in a dose-specific manner. Our results indicate that CD133(+) HSPC from umbilical cord blood may have a greater differentiation potential than previously recognized and give rise to proliferative endothelial cells participating in placental vasculogenesis.
Baal
Nelli
N
Department of Obstetrics, Justus-Liebig-University, Giessen, Germany.
Reisinger
Kerstin
K
Jahr
Henning
H
Bohle
Rainer M
RM
Liang
Olin
O
Münstedt
Karsten
K
Rao
C V
CV
Preissner
Klaus T
KT
Zygmunt
Marek T
MT
D000071916:AC133 Antigen; D015703:Antigens, CD; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics*; D042783:Endothelial Cells / Q000166:cytology; D005312:Fetal Blood / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D006023:Glycoproteins; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D018919:Neovascularization, Physiologic; D050814:Octamer Transcription Factor-3; D010455:Peptides; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics*
D000071916:AC133 Antigen; D015703:Antigens, CD; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C000606023:PROM1 protein, human; D010455:Peptides; D014157:Transcription Factors