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

Nelli Baal; Kerstin Reisinger; Henning Jahr; Rainer M. Bohle; Olin Liang; Karsten Münstedt; C.V. Rao; Klaus T. Preissner; Marek T. Zygmunt

doi:10.1160/TH04-02-0079

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2004; 92(04): 767-775
DOI: 10.1160/TH04-02-0079

Rapid and Short Communication

Schattauer GmbH

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

Nelli Baal

¹Departments of Obstetrics and Gynecology

,

Kerstin Reisinger

¹Departments of Obstetrics and Gynecology

,

Henning Jahr

³Internal Medicine and

,

Rainer M. Bohle

⁴Pathology, Justus-Liebig-University Giessen, Germany

,

Olin Liang

¹Departments of Obstetrics and Gynecology

,

Karsten Münstedt

¹Departments of Obstetrics and Gynecology

,

C.V. Rao

⁵Department of Obstetrics and Gynecology, University of Louisville, Louisville, Kentucky, USA

,

Klaus T. Preissner

,

Marek T. Zygmunt

¹Departments of Obstetrics and Gynecology

› Author Affiliations

Abstract

Summary

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.

Keywords

CD133+ cells - umbilical cord blood - Oct-4 - pluripotency

Full Text

References

References
1 Verfaillie CM. Adult stem cells: assessing the case for pluripotency. Trends Cell Biol 2002; 12: 502-8.
2 Scholer HR, Ruppert S, Suzuki N. et al. New type of POU domain in germ line-specific protein Oct-4. Nature 1990; 344: 435-9.
3 Prusa AR, Marton E, Rosner M. et al. Oct-4expressing cells in human amniotic fluid: a new source for stem cell research?. Hum Reprod 2003; 18: 1489-93.
4 Pesce M, Scholer HR. Oct-4: gatekeeper in the beginnings of mammalian development. Stem Cells 2001; 19: 271-8.
5 Ryan AK, Rosenfeld MG. POU domain family values: flexibility, partnerships, and developmental codes. Genes Dev 1997; 11: 1207-25.
6 Niwa H. Molecular mechanism to maintain stem cell renewal of ES cells. Cell Struct Funct 2001; 26: 137-48.
7 Henderson JK, Draper JS, Baillie HS. et al. Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens. Stem Cells 2002; 20: 329-37.
8 Zygmunt M, Herr F, Keller-Schoenwetter S. et al. Characterization of human chorionic gonadotropin as a novel angiogenic factor. J Clin Endocrinol Metab 2002; 87: 5290-96.
9 Liang O, Korff T, Eckhardt J. et al. Oncodevelopmental Alpha-fetoprotein (AFP) as a pro-angiogenic factor in the fetomaternal unit. J Clin Endocrinol Metab 2004; 89: 1415-22.
10 Zhang Y, Dufau ML. Silencing of transcription of the human luteinizing hormone receptor gene by histone deacetylase-mSin3A complex. J Biol Chem 2002; 277: 33431-8.
11 Fink L, Seeger W, Ermert L. et al. Real-time quantitative RT-PCR after laser-assisted cell picking. Nat Med 1998; 04: 1329-33.
12 Finke J, Fritzen R, Ternes P. et al. An improved strategy and a useful housekeeping gene for RNA analysis from formalin-fixed, paraffin-embedded tissues by PCR. Biotechniques 1993; 14: 448-53.
13 Königshoff M, Wilhelm J, Bohle RM. et al. HER-2/neu gene copy number quantified by real-time PCR: comparison of gene amplification, heterozygosity, and immunohistochemical status in breast cancer tissue. Clin Chem 2003; 49: 219-29.
14 Reshef E, Lei ZM, Rao CV. et al. The presence of gonadotropin receptors in nonpregnant human uterus, human placenta, fetal membranes, and decidua. J Clin Endocrinol Metab 1990; 70: 421-30.
15 Dufau ML. The luteinizing hormone receptor. Annu Rev Physiol 1998; 60: 461-96.
16 Forraz N, Pettengell R, Deglesne PA. et al. AC133+ umbilical cord blood progenitors demonstrate rapid self-renewal and low apoptosis. Br J Haematol 2002; 119: 516-24.
17 Kohler T, Plettig R, Wetzstein W. et al. Defining optimum conditions for the ex vivo expansion of human umbilical cord blood cells Influences of progenitor enrichment, interference with feeder layers, early-acting cytokines and agitation of culture vessels. Stem Cells 1999; 17: 19-24.
18 Yin AH, Miraglia S, Zanjani ED. et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997; 90: 5002-12.
19 Goussetis E, Theodosaki M, Paterakis G. et al. A functional hierarchy among the CD34+ hematopoietic cells based on in vitro proliferative and differentiative potential of AC133+CD34(bright) and AC133(dim/)CD34+ human cord blood cells. J Hematother Stem Cell Res 2000; 09: 827-40.
20 Shamblott MJ, Axelman J, Littlefield JW. et al. Human embryonic germ cell derivatives express a broad range of developmentally distinct markers and proliferate extensively in vitro . Proc Natl Acad Sci USA 2001; 98: 113-8.
21 Hodes RJ. Telomere length, aging, and somatic cell turnover. J Exp Med 1999; 190: 153-6.
22 Chambers I, Colby D, Robertson M. et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 2003; 113: 643-55.
23 Mitsui K, Tokuzawa Y, Itoh H. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 2003; 113: 631-42.
24 Gehling UM, Ergun S, Schumacher U. et al. In vitro differentiation of endothelial cells from AC133-positive progenitor cells. Blood 2000; 95: 3106-12.
25 Eggermann J, Kliche S, Jarmy G. et al. Endothelial progenitor cell culture and differentiation in vitro: a methodological comparison using human umbilical cord blood. Cardiovasc Res 2003; 58: 478-86.
26 Zigova T, Song S, Willing AE. et al. Human umbilical cord blood cells express neural antigens after transplantation into the developing rat brain. Cell Transplant 2002; 11: 265-74.
27 Ishikawa F, Drake CJ, Yang S. et al. Transplanted human cord blood cells give rise to hepatocytes in engrafted mice. Ann N Y Acad Sci 2003; 996: 174-85.
28 Verfaillie CM, Pera MF, Lansdorp PM. Stem cells: hype and reality. Hematology 2002; 01: 369-91.
29 Wagers AJ, Sherwood RI, Christensen JL. et al. Little evidence for developmental plasticity of adult hematopoietic stem cells. Science 2002; 297: 2256-9.
30 Pochampally RR, Smith JR, Ylostalo J. et al. Serum deprivation of human marrow stromal cells (hMSCs) selects for a sub-population of early progenitor cells with enhanced expression of Oct-4 and other embryonic genes. Blood 2004; 103: 1647-52.
31 D’Ippolito G, Diabira S, Howard GA. et al. Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential. Cell Sci 2004; 117: 2971-81.