Expression of transcription factor Oct-4 and other embryonic genes in CD133 positive cells from human umbilical cord blood

Journal: Thrombosis and Haemostasis
ISSN: 0340-6245

Paediatric Haemostasis

Issue: 2004: 92/4 (Oct) pp. 672-895
Pages: 767-775

Expression of transcription factor Oct-4 and other embryonic genes in CD133 positive cells from human umbilical cord blood

Nelli Baal 1, Kerstin Reisinger 1, Henning Jahr 3, Rainer M. Bohle 4, Olin Liang 1, 2, Karsten Münstedt 1, C.V. Rao 5, Klaus T. Preissner 2, Marek T. Zygmunt1
1 Departments of Obstetrics and Gynecology, 2 Biochemistry, 3 Internal Medicine and 4Pathology, Justus-Liebig-University Giessen, Germany 5Department of Obstetrics and Gynecology, University of Louisville, Louisville, Kentucky, USA


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 β-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.