Korean J Hematol 1997; 32(3):
Published online September 30, 1997
© The Korean Society of Hematology
냉동 제대혈 세포의 체외 증폭
김삼용, 김철희, 배광봉, 김현수, 박상준, 김종숙, 윤환중, 조덕연
충남대학교 의과대학 내과학교실
Ex vivo Expansion of Cryopreserved Cord Blood Cells
Background: Cord blood(CB), which has no HLA restriction, is an alterative to bone marrow for hematopoietic stem cell transplantation. The use of cord blood, however, is limited by the number of progenitor/stem cells necessary to reconstitute the older child or adult. Therefore, ex vivo expansion of CB could have tremendous impact on diverse clinical settings. We studied the ex vivo expansion of isolated population of CD34 cells from cryopreserved CB cells.
Methods: CD34 cells were isolated from cryopresered CB mononuclear cells. Purified cells were cultured with various combinations of hematopoietic growth factors including erythropoietin(EPO),stem cellfactor(SCF), granulocyte-colony-stimulating factor(G-CSF), granulocyte, macrophage-colony-stimulatingfactor(GM-CSF), interleukin-1 β (IL-1 β ), IL-3, and IL-6. After 7, 10 or 14 days of culture, the fold increases of colony-forming unit- granulocyte, macrophage(CFU-GM), burst-forming
unit-erythroid(BFU-I), colony-forming unit-mix(CFU-Mix), and high proliferative potential colony-forming cell(HPP-CFC) were evaluated.
Results: Ten-day culture with the combination of EPO, SCF, G-CSF, IL-1 β , and IL-3 resulted in a median of 60-fold increase of CFU-GM, which was greater than those with the combinations of less than 5 growth factors. The addition of IL-6 or GM-CSF to this combination did not enhance CFU-GM expansion. Ten-day culture was significantly superior to 7-day culture for CFU-GM expansion. Prolongation of culture to 14 days, however, revealed decreased expansion of CFU-GM compared to 10 days.
CFU-E and CFU-Mix were expanded to 2∼5 folds in 7-day culture with the combination of EPO, SCF, and G-CSF. Further expansion was not achieved in 10-day culture and colonies disappeared in 14-day culture. HPP-CFC was expanded to a median of 7.5 folds in 7-day culture with the combination of EPO, SCF, G-CSF, IL-1 β , IL-3,
and IL-6. Neither 10-day or 14 day-culture enhanced expansion of HPP-CFU.
Conclusion: Cryopresered cord blood cells maintain ex vivo expansion potential. In our system, 10-day culture with the combination consisting of EPO, SCF, G-CSF, IL-1
β , and IL-3 seems to be adequate for hematopoietic progenitor/stem cell expansion from cryopreserved cord blood cells.
Keywords Cord blood cells; Hematopoietic growth factors; Ex vivo expansion;
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Korean J Hematol 1997; 32(3): 347-359
Published online September 30, 1997
Copyright © The Korean Society of Hematology.
냉동 제대혈 세포의 체외 증폭
김삼용, 김철희, 배광봉, 김현수, 박상준, 김종숙, 윤환중, 조덕연
충남대학교 의과대학 내과학교실
Ex vivo Expansion of Cryopreserved Cord Blood Cells
Samyong Kim, Chu Hee Kim, Gwang Bong Bae, Hyun Soo Kim, Sang Jun Park, Jong Suk Kim, Hwan Jung Yun, Deog Yeon Jo
Department of Internal Medicine, Chungnam National University College of Medicine, Taejon, Korea
Abstract
Background: Cord blood(CB), which has no HLA restriction, is an alterative to bone marrow for hematopoietic stem cell transplantation. The use of cord blood, however, is limited by the number of progenitor/stem cells necessary to reconstitute the older child or adult. Therefore, ex vivo expansion of CB could have tremendous impact on diverse clinical settings. We studied the ex vivo expansion of isolated population of CD34 cells from cryopreserved CB cells.
Methods: CD34 cells were isolated from cryopresered CB mononuclear cells. Purified cells were cultured with various combinations of hematopoietic growth factors including erythropoietin(EPO),stem cellfactor(SCF), granulocyte-colony-stimulating factor(G-CSF), granulocyte, macrophage-colony-stimulatingfactor(GM-CSF), interleukin-1 β (IL-1 β ), IL-3, and IL-6. After 7, 10 or 14 days of culture, the fold increases of colony-forming unit- granulocyte, macrophage(CFU-GM), burst-forming
unit-erythroid(BFU-I), colony-forming unit-mix(CFU-Mix), and high proliferative potential colony-forming cell(HPP-CFC) were evaluated.
Results: Ten-day culture with the combination of EPO, SCF, G-CSF, IL-1 β , and IL-3 resulted in a median of 60-fold increase of CFU-GM, which was greater than those with the combinations of less than 5 growth factors. The addition of IL-6 or GM-CSF to this combination did not enhance CFU-GM expansion. Ten-day culture was significantly superior to 7-day culture for CFU-GM expansion. Prolongation of culture to 14 days, however, revealed decreased expansion of CFU-GM compared to 10 days.
CFU-E and CFU-Mix were expanded to 2∼5 folds in 7-day culture with the combination of EPO, SCF, and G-CSF. Further expansion was not achieved in 10-day culture and colonies disappeared in 14-day culture. HPP-CFC was expanded to a median of 7.5 folds in 7-day culture with the combination of EPO, SCF, G-CSF, IL-1 β , IL-3,
and IL-6. Neither 10-day or 14 day-culture enhanced expansion of HPP-CFU.
Conclusion: Cryopresered cord blood cells maintain ex vivo expansion potential. In our system, 10-day culture with the combination consisting of EPO, SCF, G-CSF, IL-1
β , and IL-3 seems to be adequate for hematopoietic progenitor/stem cell expansion from cryopreserved cord blood cells.
Keywords: Cord blood cells, Hematopoietic growth factors, Ex vivo expansion,
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