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KMID : 0653520000050010069
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Korean Journal of Hematopoietic Stem Cell Transplanstation
2000 Volume.5 No. 1 p.69 ~ p.79
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Ex Vivo Expansion of Myeloid Progenitors by Using Hematopoietic Growth Factors
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Sung Ki-Woong
Lee Yun-Hee
Jung Hye-Lim
Koo Hong-Hoe
Shin Hee-Young
Ahn Hyo-Seop
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Abstract
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Background : If myeloid progenitors expanded ex vivo are transplanted as adjuvant to unprocessed cells, neutropenic period after stem cell transplantation will be shortened without affecting long-term engraftment. Unlike previous studies which have focused primarilly on expansion of primitive hematopoietic stem cells with controversial results, in this study, ex vivo expansion of myeloid progenitors was primarilly focused on. For maximal expansion of myeloid progenitors, optimal combination of hematopoietic growth factors, optimal duration of culture, and difference in ability of expansion according to source of hematopoietic cells were studied.
Methods : CD34+ cells isolated from cord blood (CB) and mobilized peripheral blood (PB) were cultured and ex vivo expanded into myeloid progenitors by using hematotopoietic growth factors (IL 1-β, IL-3, IL-6, SCF, G-CSF, GM-CSF) which are known to stimulate differentiation and proliferation of myeloid progenitors. To find optimal combination of growth factors, CD34+ cells isolated from CB were cultured for 9 days with various combinations of growth factors. To find optimal duration of culture and difference in ability of expansion, CD34+ cells isolated from CB and PB were cultured for 7, 9, 12, 14, 21, 28 days.
Results : Myeloid progenitors were successfully produced from CD34+ cells. Maximal expansion was obtained with the combination of IL-3+SCF+G-CSF+GM-CSF. SCF was thought to be most important growth factor for expansion of myeloid progenitors. Optimal duration of culture was different according to sources of CD34+ cells. CD34+ cells obtained from CB were more easily expanded into myeloid rogenitors than PB, but optimal duration of culture in CB (12 days) was longer than PB (9 days). After reaching maximal expansion of progenitors, number of nucleated cell increased, but number of myeloid progenitors decreased. After 28 days of culture, nearly all viable cells were mature myeloid cells. IL 1-β and IL-6 did not show any additive or synergistic effect on expansion of myeloid progenitors.
Conclusion : CD34+ cells can be successfully expanded into myeloid progenitors by using hematopoietic growth factors. Optimal condition of culture was different according to sources of CD34+ cells. Neutropenic period after stem cell transplantation will be shortened by using ex vivo expanded myeloid
progenitors.
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KEYWORD
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Ex vivo expansion, Transplantation, Myeloid progenitor
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