2004 Fiscal Year Final Research Report Summary
Application of ex vivo expanded hematopoietic stem cells stimulated with human umbilical venous endothelial cells for transplantation in children
| Project/Area Number |
15591115
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Saga University (2004)
佐賀医科大学 (2003) |
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Principal Investigator |
FUJITA Ichiro Saga University, Pediatrics, Assistant Professor, 医学部, 講師 (60228989)
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| Co-Investigator(Kenkyū-buntansha) |
ISHII Eiichi Saga University, Pediatrics, Associate Professor, 助教授 (20176126)
NAKAHATA Tatsutoshi Kyoto University, Pediatrics, Professor, 医学研究科, 教授 (20110744)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Endothelial cells / Hematopoietic stem cells / Co-culture system / Fluorescent immunocytochemistory / Inflammation-related proteins / Oxidized LDL / IFN-γ |
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| Research Abstract |
The aim of this study was to establish the ex vivo expansion of hematopoietic stem cells by co-culture system with human umbilical venous endothelial cells(HUVEC). After the establishment of the co-culture system with HUVEC, hematopoietic stem cells continuously grew and proliferated. However, several problems including how to maintain the function of endothelial cells for a long time, or how to make an attachment of stem cells effectively with endothelial cells via membrane filter.
The aim of the next study was to clarify effects of INF-γ on monocytes and macrophages, using U937 monocytic cells. We investigated expressions of inflammation-related proteins such as MCP-1 (Monocyte chemoattractant protein-1,COX-2 (Cyclooxyganase-2), iNOS (inducible nitric oxide synthetase), Ferritin, CRP(C-reactive protein), AngiotensinII receptor-1(AT1R), Scavenger receptor type A(SRA) and Oxidized LDL receptor-1(LOX-1) with fluorescent immnocytochemical staining. Stimulation with INF-γ for 24 hours caused upregulation of all these proteins in U937 cells strongly. The fluorescent intensity of these proteins was increased by more than 150% of that in unstimulated cells. In comparison with TNF-α, which is one of proinflammatory cytokines, TNF-α did not give such striking effect on U937 cells. Moreover, INF-γ seemed to accelerate uptake of Dil labeled oxidized LDL and induced morphological changes like foam cells in some cells. These results demonstrate that IFN-γ could activate monocytes strongly and variously, and that effects of FN-γ could play important roles in disease such as hemophagocytic syndrome, macrophage activating syndrome and atherosclerosis.
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