Source:http://linkedlifedata.com/resource/pubmed/id/11861065
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1-2
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pubmed:dateCreated |
2002-3-7
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pubmed:abstractText |
The object of this study is to explore a culture method to generate a large number of functional and mature dendritic cells (DC) from human CD34+ hematopoietic progenitor cells. In the present study, we used a two-step method combined with calcium ionophore to induce DC from cord blood (CB) or normal human bone marrow (BM) CD34+ progenitor cells. The two-step method consists of 10 days of first step culture for the expansion and proliferation of CD34+ hematopoietic progenitor cells in the presence of SCF, IL-3, IL-6, G-CSF, and 7--11 days of second step culture for the induction of DC in the presence of GM-CSF, IL-4 and TNF-alpha. By the two-step culture, total nucleated cells were increased 208+/-66 (+/-SD, n=13), or 94+/-29 (n=5)-fold in the culture of CB or BM cells, respectively, compared with the number of CD34+ cells at the time of starting culture. Out of the total nucleated cells, 23 +/-10.4% of cells in CB cell culture and 25 +/-5% of cells in the BM cell culture acquired DC characteristic phenotypes, which were marked expressions of CD1a, HLA-DR, co-stimulatory molecules such as CD80, CD40, and adhesion molecule such as CD58. In allogeneic mixed leukocyte reaction (MLR), two-step cultured cells showed potent allo-stimulatory capacity. With this two-step culture, the absolute number of CD1a+ cells that co-expressed HLA-DR, CD80, CD40 and CD58 was enhanced approximately 3 times in CB cell culture and 1.9 times in BM cell culture, compared with the commonly used one-step culture method for the generation of DC from CD34+ cells using SCF, GM-CSF and TNF-alpha. However, on these DC generated in the two-step culture, the expressions of co-stimulatory molecule CD86 and mature DC marker CD83 were not sufficient. By the treatment of two-step cultured cells with calcium ionophore agent (A23187), the expression of co-stimulatory molecules such as CD86 and CD80 (especially CD86) was up-regulated. Besides, the expression of mature DC marker CD83 was remarkably induced by treatment with A23187 for a short duration (24 h). Consistent with the up-regulation of surface molecules CD86, CD80 and CD83, the two-step cultured cells treated with A23187 also showed a stronger allo-stimulatory capacity compared with the cells without A23187 treatment. In conclusion, the present study demonstrated that the two-step culture method effectively improved the yield of CD1a+ DC generated from CD34+ cells, and the phenotypes and functions of these CD1a+ DC could be enhanced efficiently by treatment with a calcium ionophore agent.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0022-1759
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pubmed:author |
pubmed-author:AbeTakashiT,
pubmed-author:AizawaYoshifusaY,
pubmed-author:FurukawaTatsuoT,
pubmed-author:FuseIchiroI,
pubmed-author:KanazawaNaokoN,
pubmed-author:LiuAichunA,
pubmed-author:NaritaMiwakoM,
pubmed-author:NikkuniKohjiK,
pubmed-author:TakahashiMasuhiroM,
pubmed-author:TurkG MGM,
pubmed-author:ZhengZhiyinZ
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
261
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
49-63
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:11861065-Antigens, CD34,
pubmed-meshheading:11861065-Bone Marrow Cells,
pubmed-meshheading:11861065-Calcimycin,
pubmed-meshheading:11861065-Cell Culture Techniques,
pubmed-meshheading:11861065-Cell Differentiation,
pubmed-meshheading:11861065-Dendritic Cells,
pubmed-meshheading:11861065-Fetal Blood,
pubmed-meshheading:11861065-Humans,
pubmed-meshheading:11861065-Ionophores
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pubmed:year |
2002
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pubmed:articleTitle |
Generation of functional and mature dendritic cells from cord blood and bone marrow CD34+ cells by two-step culture combined with calcium ionophore treatment.
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pubmed:affiliation |
First Department of Internal Medicine, Faculty of Medicine, Niigata University, Niigata, Japan.
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pubmed:publicationType |
Journal Article,
In Vitro
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