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In vitro generation of human CD141+ dendritic cells from CD14+ monocytes

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dc.contributor.advisor윤철희-
dc.contributor.author김수지-
dc.date.accessioned2017-10-31T07:51:47Z-
dc.date.available2018-01-23-
dc.date.issued2017-08-
dc.identifier.other000000145790-
dc.identifier.urihttps://hdl.handle.net/10371/137582-
dc.description학위논문 (석사)-- 서울대학교 대학원 농업생명과학대학 농생명공학부, 2017. 8. 윤철희.-
dc.description.abstractHuman CD141+ (BDCA-3+) dendritic cells (DCs) are specialized to cross-presentation and, thus extensively studied for developing DC-based therapy against cancer. A series of attempts was made to generate CD141+ DCs from cord blood CD34+ hematopoietic progenitors to overcome a practical limitation, in vivo rareness. However, in vitro differentiation of CD141+ DC needs to be further investigated. In the present study, I identified CD141 expression in the well-known culture system that generates DCs from CD14+ monocytes in presence of GM-CSF and IL-4. After 8-days of the culture, CD141 was detected only on the cells which adhered to the bottom of the culture plate. The attached cells exhibited a typical feature of immature monocyte-derived DCs (moDCs), except for higher CD86 expression, more dendrites, and higher granularity compared to those which did not attach. Additional 3 days of culture further increased CD141 expression in the cells retaining adhesion, which partially expressed CELC9A. Indeed, they exhibited relatively effective uptake of dead cells. Interestingly, the attached moDCs differently responded to polyinosinic:polycytidylic acid (poly I:C) stimulation as well as the mixed lymphocyte reaction. Collectively, in vitro generation of human CD141+ DCs from CD14+ monocytes will contribute to further investigation into yielding human CD141+ DCs using peripheral blood monocytes.-
dc.description.tableofcontentsI. Review of Literature 1
1. Human monocyte-derived DC 1
1.1 Monocyte 1
1.2 In vitro derived moDCs 2
2. Human CD141+ dendritic cell 3
2.1 Introduction 3
2.2 Characterization of CD141+ DCs 5
2.3 The role of CD141+ DCs in T cell responses 6
2.4 Cross-presentation of CD141+ DCs 7
2.5 Development of CD141+ DCs 8
II. Introduction 9
III. Materials and Methods 12
1) Ethical statement 12
2) Isolation of human monocytes 12
3) CD14+ monocyte culture 13
4) WrightGiemsa Staining 13
5) Flow cytometry 14
6) PolyI:C stimulation 14
7) Mixed lymphocyte reaction 15
8) Dead cell uptake 15
9) Enzyme-linked immunosorbent assay (ELISA) 15
10) Statistical analysis 16
IV. Results 17
1) Three different forms of moDCs after the differentiation of peripheral blood CD14+ monocytes 20
2) Attached cells after CD14+ monocyte culture are immature DCs with relatively high expression of CD141 and CD86 23
3) Non-attached cells are not important to CD141 expression of the attached cells in GM-CSF/IL-4-driven culture system 26
4) Adhesive capacity is partially involved in CD141 expression of moDCs 28
5) Dead cells uptake and TLR3 response of Attached CD141+ moDCs 31
6) Allogeneic mixed lymphocyte reaction by attached CD141+ moDCs 34
V. Supplementary results 35
1) The absolute number of nine different subsets after additional 3-day culture 35
2) Surface molecules of nine different subpopulations after additional 3-day culture 36
VI. Discussion 37
VII. Literature Cited 40
VIII. Summary in Korean 47
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dc.formatapplication/pdf-
dc.format.extent1189679 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectCD141-
dc.subjectMonocyte-derived dendritic cells-
dc.subjectAdhesion-
dc.subjectGM-CSF/IL-4-driven culture system-
dc.subject.ddc630-
dc.titleIn vitro generation of human CD141+ dendritic cells from CD14+ monocytes-
dc.typeThesis-
dc.description.degreeMaster-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2017-08-
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