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ctt-journal > Bilko et al. (Abstract)

Bilko et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract16

© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported

Abstract accepted for "5th Raisa Gorbacheva Memorial Meeting Hematopoietic Stem Cell Transplantation in Children and Adults", Saint Petersburg, Russia, September 18–20, 2011

Preliminary Program

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Expansion of AC133+ cells from cord blood in long-term culture in vitro

Nadiia M. Bilko, Iryna Z. Borbulyak, Denys I. Bilko

National University “Kyiv-Mohyla Academy”, Kiev, Ukraine

Correspondence: Nadiia M. Bilko, National University “Kyiv-Mohyla Academy”, 2, Skovoroda str., Kiev, 04655, Ukraine, E-mail: nbilko@spam is badukma.kiev.ua

Abstract

One of the most important goals in cell biotechnology is the determination of the conditions that will allow the expansion of hematopoietic progenitor cells. The aim of the current investigations was to establish a model for the determination of conditions for expansion of AC133+ cells separated from human cord blood.

An original model of gel diffusion chambers (DC) was implemented; purified cells were injected into the inner cavity of the DC in DMEM culture medium with 15% FCS, and DC were submerged in 6-well plates supplemented with DMEM, 15% FCS, L-glutamine, 0.1 mM β-mercaptoethanol, growth factors in different combinations (1: Flt-3, SCF, IL-3; 2: IL-6, GM-CSF; and 3: SCF, IL-3, GM-CSF) and cultivated over 5 weeks. In the last two cases cultivated cells were pre-incubated with Flt-3.

In previous investigations we showed that this treatment before cultivation promotes primitive AC133+ cells to further stages of development and increases the sensitivity of hematopoietic cells to cytokines. Following cultivation in vitro in the presence of the third combination of cytokines with pre-incubation with Flt-3, this combination provided long-term maintenance of hematopoiesis ex vivo and 17-fold expansion. This model can be used for determination of appropriate conditions for stem cells and progenitor cells expansion in vitro.

Keywords: stem cells, hematopoietic progenitor cells, growth promoting cytokines, ex vivo cultural system