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KMID : 1094720100150010157
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 1 p.157 ~ p.166
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Ex vivo Expansion of Hematopoietic Cells from CD34+ Cord Blood Cells in Various Culture Conditions
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Choi Yong-Woon
Park Hyun-Hee
Oh Duk-Jae
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Abstract
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This study compared the cell expansion and colony-forming ability of human cord blood stem cells cultured ex vivo with 2 types of cytokine combinations, and 2 types of media characterized by the presence or absence of fetal bovine serum (FBS) in 2 or 3 dimensional (2D or 3D) culture environments. Purified CD34+ cells derived from different donors were cultured in Iscove뭩 Modified Dulbecco뭩 Medium (IMDM) and Ultraculture serum-free medium (SFM) containing the cytokine cocktail-I (coc-I) (EPO, GM-CSF, SCF, and IL-3) or cytokine cocktail-II (coc-II) (TPO, G-CSF, SCF, IL-6, and Flt3/Flk-2 ligand) with or without FBS. Generally, higher CFU-GM values were observed in the IMDM compared to the SFM. In the coc-I conditions, the 멗MDM + coc-I?and 멗MDM + coc-I + FBS?conditions gave the greatest cell (1,667 ?274 and 1,600 ?140-fold, respectively) and colony-forming units (CFU) expansions (BFU-E: 21 ?3, 36 ?5; CFU-GM: 95 ?19, 81 ?17; and CFU-GEMM: 2 ?1, 3 ?1-fold, respectively) in 26 day culture, respectively. In the coc-II conditions, the 멣FM + coc-II?condition gave the greatest cell expansion (2,143 ?134-fold), but the 멗MDM + coc-II?condition gave the best CFU-GM expansion (924 ?110-fold) in 26 day culture. In conclusion, 멗MDM + coc-I? and 멗MDM + coc-II?were the most accessible conditions for CFU expansion for all culture cases. The 2D stationary culture had affirmative effect on CFU expansion compared to the 3D culture using semisolid MethocultTM. These results are believed to be significant in the ��=翟蹟 expansion of hematopoietic stem cells.
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KEYWORD
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ex vivo expansion, cord blood, CD34+ cell
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