In vitro expansion and lineage commitment of the human umbilical cord blood myeloid progenitors
- Authors: Petyovka NV1, Goncharova NV1, Seviaryn IM1, Kosmacheva SM1, Potapnev MP1
-
Affiliations:
- Belarusian Research & Production Center for Hematology & Transfusiology
- Issue: Vol 7, No 1 (2012)
- Pages: 40-48
- Section: Articles
- URL: https://genescells.ru/2313-1829/article/view/121680
- DOI: https://doi.org/10.23868/gc121680
- ID: 121680
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Abstract
In vitro proliferation and myeloid differentiation of human
umbilical cord blood (UCB) CD34+ cells with/without bone
marrow multipotential mesenchymal stromal cells (BMMMSCs)
feeding layer in serum-free medium supplemented
with IL-3, IL-6, SCF and Flt3/l were investigated.
Mononuclear cells (MCs) were isolated from the healthy
donor UCB specimens (n = 4), CD34-enriched MCs were
obtained by positive immunomagnetic selection (CD34+-rich).
Both cell fractions (MCs and CD34+-rich) were expanded for
two weeks in vitro. Cells were seeded to the new vessels
when necessary. The percentage of CD34/CD45-positive cells
was accounted by flow cytometry, subpopulations of myeloid
progenitors were assessed in colony-forming tests.
Upon two weeks of culture the number of CD34+ cells in
MC fraction augmented by 70±29 and 980±414-fold with
and without BM-MMSC feeding layer respectively and was
not significantly differed from that in CD34+-rich fraction.
Progenitor cell subpopulations were not altered during the
first week of culture but granulocytopoiesis was superior
over erythropoiesis during the second week. CD34 and
CD45 expression patterns revealed two hematopoietic cell
populations, one of them differentiating on the first week of
expansion, and the other - on the second one. BM-MMSC
feeder promoted significantly higher (p < 0,05) expansion
rates of CD34+ cells and myeloid progenitors in comparison
with non-feeding culture. BM-MMSC feeder also caused
progenitor cells redistribution between suspension and
adhesive fractions by predominantly binding erythroid and
multipotential progenitors. Optimum expansion protocol for
multipotential progenitors was culturing CD34+-rich cell
population with BM-MMSC feeding layer for a week which
resulted in the increase the amount of the nucleated cells,
CD34+ cells, multi-potential progenitors, and colony-forming
units by 56, 36, 45, and 60-fold respectively.
umbilical cord blood (UCB) CD34+ cells with/without bone
marrow multipotential mesenchymal stromal cells (BMMMSCs)
feeding layer in serum-free medium supplemented
with IL-3, IL-6, SCF and Flt3/l were investigated.
Mononuclear cells (MCs) were isolated from the healthy
donor UCB specimens (n = 4), CD34-enriched MCs were
obtained by positive immunomagnetic selection (CD34+-rich).
Both cell fractions (MCs and CD34+-rich) were expanded for
two weeks in vitro. Cells were seeded to the new vessels
when necessary. The percentage of CD34/CD45-positive cells
was accounted by flow cytometry, subpopulations of myeloid
progenitors were assessed in colony-forming tests.
Upon two weeks of culture the number of CD34+ cells in
MC fraction augmented by 70±29 and 980±414-fold with
and without BM-MMSC feeding layer respectively and was
not significantly differed from that in CD34+-rich fraction.
Progenitor cell subpopulations were not altered during the
first week of culture but granulocytopoiesis was superior
over erythropoiesis during the second week. CD34 and
CD45 expression patterns revealed two hematopoietic cell
populations, one of them differentiating on the first week of
expansion, and the other - on the second one. BM-MMSC
feeder promoted significantly higher (p < 0,05) expansion
rates of CD34+ cells and myeloid progenitors in comparison
with non-feeding culture. BM-MMSC feeder also caused
progenitor cells redistribution between suspension and
adhesive fractions by predominantly binding erythroid and
multipotential progenitors. Optimum expansion protocol for
multipotential progenitors was culturing CD34+-rich cell
population with BM-MMSC feeding layer for a week which
resulted in the increase the amount of the nucleated cells,
CD34+ cells, multi-potential progenitors, and colony-forming
units by 56, 36, 45, and 60-fold respectively.
About the authors
N V Petyovka
Belarusian Research & Production Center for Hematology & TransfusiologyBelarusian Research & Production Center for Hematology & Transfusiology
N V Goncharova
Belarusian Research & Production Center for Hematology & TransfusiologyBelarusian Research & Production Center for Hematology & Transfusiology
I M Seviaryn
Belarusian Research & Production Center for Hematology & TransfusiologyBelarusian Research & Production Center for Hematology & Transfusiology
S M Kosmacheva
Belarusian Research & Production Center for Hematology & TransfusiologyBelarusian Research & Production Center for Hematology & Transfusiology
M P Potapnev
Belarusian Research & Production Center for Hematology & TransfusiologyBelarusian Research & Production Center for Hematology & Transfusiology
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