Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Proliferation, Apoptosis, and Telomerase Activity in Human Cord Blood CD34+ Cells Cultured with Combinations of Various Cytokines

  • Ahn, Myung-Ju (Department of Internal Medicine, Hanyang University, College of School) ;
  • Lee, Hye-Sook (Department of Internal Medicine, Hanyang University, College of School) ;
  • Jang, Mi-Yune (Biochemistry, Hanyang University, College of School) ;
  • Choi, Jung-Hye (Department of Internal Medicine, Hanyang University, College of School) ;
  • Lee, Young-Yeul (Department of Internal Medicine, Hanyang University, College of School) ;
  • Park, Hyung-Bae (Department of Internal Medicine, Hanyang University, College of School) ;
  • Lee, Yong-Sung (Biochemistry, Hanyang University, College of School)
  • Published : 2003.06.01
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Abstract

Umbilical cord blood (UCB), a rich source of hematopoietic stem/progenitor cells, has been proposed as an alternative to bone marrow and peripheral blood for transplantation treatment. Ex vivo expansion of cord blood stem cells could make the use of cord blood transplant feasible even for adult patients. However, the optimal cytokine cocktail for expansion of stem cells is yet to be established. This study compares proliferation, apoptosis, and telomerase activities in human cord blood stem cells cultured ex vivo with FLT3 ligand (FL)/thrombopoietin (TPO) or FL/TPO/stem cell factor (SCF), with a view to determine optimal combination of cytokines. CD34+ cells were cultured in DMEM containing either FL (50 ng/ml) and TPO (10 ng/ml) (FT group) or FL (50 ng/ml), TPO (10 ng/ml) and SCF (50 ng/ml) (FTS group). The cell proliferation rate was ten times higher in the FTS group. Although cells cultured with the two different combinations of cytokines were maintained for a long term (up to 8 weeks), a large number of cells underwent differentiation during this period. Cells cultured in FTS displayed lower levels of apoptosis compared to those of the FT group during the Initial 7 days of culture. The CD34+ fraction in both groups was markedly decreased to $21-30\%$ , and only $5-6\%$ was detected at 14 days of culture. Telomerase activity detected in human CD34+ cord blood at low levels was upregulated during the early phase of culture and decreased to baseline levels in the later phase. The telomerase activity of cord blood cultured in FT was lower than that of the FTS group. Our results suggest that, on adding stem cell factors to the FT cytokines, cultured CD34+ cord blood cells display a greater degree of cell proliferation and decreased apoptosis. However, during CD34+ cord blood cell culture, a Barge number of cells undergo differentiation, indicating that more potent novel cytokines or new culture conditioning methods should be developed to maintain their ability to engraft and sustain long-term hematopoiesis.

Keywords

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