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Bilko et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 9
doi: 10.3205/ctt-2010-No9-abstract15
© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported

Abstract accepted for "4th Raisa Gorbacheva Memorial Meeting on Hematopoietic Stem Cell Transplantation",
Saint Petersburg, Russia, September 18–20, 2010

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The role of Flt-3 in the enrichment of an AC133-positive cell population by hemopoietic progenitor cells in vitro

Nadiya M. Bilko, Svitlana V. Vasylovska, Denys I. Bilko, Iryna Z. Borbulyak, Alex A. Barash

National University “Kiev-Mogilanskaya Academy”, Kiev, Ukraine

Correspondence: Nadezhda M. Bilko, Professor, Chair of Biology, Faculty of Natural Sciences; Director, Center of Molecular and Cellular Studies, National University "Kiev-Mogilanskaya Academy", G. Skovoroda St., 2, 04655, Kiev, Ukraine, E-mail: nbilko@spam is badukma.kiev.ua

Abstract

Among the concept approaches used for the analysis of proliferation and differentiation of human hemopoietic progenitor cells, one is based on ex vivo cultural investigations.

The aim of this work was to investigate the clonogenic potential of an AC133-positive cell population derived from cord blood, and to evaluate the influence of incubation with Flt-3 growth factor on their functional activity in vitro.

Methods: Cord blood mononuclear cells were obtained using Histopaque density gradient centrifugation; AC133-positive cells were selected with Miltenyi Biotech system. Incubation with Flt-3 growth factor (10ng/ml) was carried out at 37 °C at 12, 16, and 18 hours before cultivation. The cells were then cultivated in suspension culture with IL-3 and GM-CSF over 3 weeks with further clonogenic assay in semisolid agar in vitro culture (CFU-GM).

Results: It was established that a 16-hour incubation of an AC133-positive cell fraction with Flt-3 growth factor promotes their enrichment with functionally active hemopoietic stem cells and progenitors upon in vitro culture, which is reflected in the increase of colony-forming activity.

The Flt-3 receptor, or CD135, refers to the family of growth factor receptors that are implicated in early embryogenesis. Flt-3 growth factor facilitates the proliferation and differentiation of early hemopoietic progenitor cells. Thus we can assume that the role of Flt-3 is to promote the primitive AC133-positive cell population to further stages of development. It increases the sensitivity of hemopoietic cells to cytokine action. As a result, we observed an expansion of AC133-positive cell numbers in cell suspension, as well as the increase in their cloning efficiency.

Keywords: hemopoietic progenitor cells, AC133-positive cord blood-derived cells, Flt-3 growth factor, clonogenic assay in vitro

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