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ctt-journal > Petyovka et al. (Abstract)

Petyovka et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract04

© 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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In vitro expansion and lineage commitment of the human umbilical cord blood myeloid progenitors

Natalie V. Petyovka, Natalia V. Goncharova, Ihar M. Seviaryn,  Svetlana M. Kosmacheva, Mikhail P. Potapnev

Belarusian Research and Production Center for Hematology and Transfusiology, Minsk, Republic of Belarus

Correspondence: Natalie V. Petyovka, Belarusian Research and Production Center for Hematology and Transfusiology, Dolginovski Trakt, 160, Minsk, Republic of Belarus, E-mail: npet@spam is badbcht.by

Abstract

Aim: The investigation of in vitro proliferation and myeloid differentiation of human umbilical cord blood (UCB) CD34+ cells with/without a bone marrow multipotential mesenchymal stromal cell (BM-MMSC) feeding layer in serum-free medium supplemented with IL-3, IL-6, SCF, and Flt3/l.

Methods: Mononuclear cells (MCs) were isolated from UCB after normal delivery (n=4), and part of the MCs were CD34-enriched by positive immunomagnetic selection (CD34+-rich). MCs and CD34+-rich cell fractions were expanded for two weeks in vitro. The cells were seeded to new vessels when necessary. The percentage of CD34/CD45-positive cells was counted by means of flow cytometry, and subpopulations of myeloid progenitors were assessed by a colony-forming test.

Results: At two weeks of culture the number of CD34+ cells in the MC fraction increased by 70±29- and 980±414-fold with and without the BM-MMSC feeding layer respectively, and were not significantly different from those in the CD34+-rich fraction. Progenitor subpopulations did not alter during the first week of culture, but granulocytopoiesis prevailed over erythropoiesis during the second week. The BM-MMSC feeder promoted significantly higher (p<0.05) expansion rates of CD34+ cells and myeloid progenitors in comparison with non-feeding culture. The BM-MMSC feeder layer also caused the progenitor cells' redistribution between suspension and adhesive fractions by predominantly binding erythroid and multipotential progenitors.

Conclusion: The optimum expansion protocol for multipotential progenitors was culturing CD34+-rich cell population with BM-MMSC feeding layer for no more than a week, which resulted in the increase of the nucleated cells, CD34+ cells, multi-potential progenitors, and colony-forming units by 56-,  36-, 45-, and 60-fold respectively.

Keywords: umbilical cord blood, hematopoietic stem cells, HSCs, СD34+ cells, progenitors, expansion