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

Authors: Nelli, Baal; Kerstin, Reisinger; Henning, Jahr; Rainer M, Bohle; Olin, Liang; Karsten, Münstedt; C V, Rao; +2 Authors

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

Abstract

SummaryA 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.

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Keywords

Pluripotent Stem Cells, Cell Culture Techniques, Endothelial Cells, Gene Expression Regulation, Developmental, Neovascularization, Physiologic, Cell Differentiation, Fetal Blood, Hematopoietic Stem Cells, DNA-Binding Proteins, Antigens, CD, Humans, AC133 Antigen, Peptides, Octamer Transcription Factor-3, Cells, Cultured, Cell Proliferation, Glycoproteins, Transcription Factors

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Powered by OpenAIRE graph
citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
63
Average
Top 10%
Top 10%