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ctt-journal > Tolmachev et al. (Abstract)

Tolmachev et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract34

© 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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Phenotypic and functional characteristics of cord blood mononuclear cells

Vladimir S. Tolmachev, Elyzaveta I. Borovitskaya, Gulsum K. Smagulova

Scientific center of obstetrics, gynecology and perinatology MH, Almaty, Kazakhstan

Correspondence: Vladimir S. Tolmachev, Scientific center of obstetrics, gynecology and perinatology MH, 125, Dostyk str., Almaty, 050020, Kazakhstan, E-mail: lab_cyto_kz@spam is badmail.ru

Abstract

Aim: To analyze the phenotypical and functional characteristics of cord blood (CB) mononuclear cells (MNC) and methods of their ex vivo expansion.

Methods: Flow cytometry (FC) analysis, CB MNC culture with IL3, IL6 and nonhistone proteins, karyotyping.

Results:
The analyzed CB samples contained up to 9.8% MNC express hematopoietic stem cell (HSC) markers (СD34+, СD38+, СD117+) and 3.2% mesenchymal stem cells (СD90+, СD73+). It was revealed that 97.9±0.9 % CB MNC are in the G0-G1 stages of a cell cycle. The MNC number increased in cultures with a placenta extract (to 30%) and IL3, IL6. The CD34+ cell number increased 2-fold and 2.5-fold in RPMI and MEM cultures with growth factors respectively. MNC quantity increased 1.3–3.5-fold in nonhistone proteins containing cultures. MNC had high viability (98 %) and low apoptotic activity (2% in a cellular cycle and 1% CD95+cells). The accumulation of ionized calcium (Ca2 +- DNA) in the MNC culture (3-fold) reflected the accelerated ageing of cells in vitro. Chromatid type aberrations (50%), stable chromosomal aberrations (29%) and an aberration of chromosomal type (12%) were noted. Acridin orange DNA staining revealed chromatin condensation.

Conclusion: The results of the current study could improve CB graft quality for clinical transplantation.

Keywords: mononuclear cells, chromosomal aberrations, DNA fluorescence