Functional and Phenotypic Characterizations of Swine Dendritic Cells Derived from Bone Marrow Haematopoietic Cells in vitro.

• Main Authors: Yu-Zhang Chen, 陳昱彰
• Other Authors: Hso-Chi Chaung
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/29007370916250109664

碩士 === 國立屏東科技大學 === 生物科技研究所 === 93 === Dendritic cells (DC) have been referred as the most professional antigen-presenting-cell, which plays important roles in modulating immune responses. In this study, bone marrow haematopoietic cells (BMHC) were isolated from the long bones of piglets and then stimulated by co-culturing with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin-4 (IL-4) to generate BMHC-derived immature DC (BMHC-imDC). Then, TNF-α was used to induce the maturation of BMHC-imDC into BMHC-derived mature DC (BMHC-mDC). Cell morphology of BMHC, BMHC-imDC and BMHC-mDC was examined by transmission electron microscopy and light microscopy. The immuno-phenotypic characteristics of these cells were analyzed by using flow cytometry. The results showed that BMHC-imDC derived from BMHC were characterized with consistent cell morphology and 85.7±1.5％ of BMHC-imDC had manifest dendritic pseudopods. Their cell surface markers were identified as SWC1med SWC3high SWC9low CD4low CD8low/med CD14med/high CD80/86low /med MHC Imed/high MHC IIlow. BMHC-mDC had no significant changes on their pseudopods during the maturation. As compared to those of BMHC-imDC, the cell percentages of BMHC-mDC with MHC II＋ and CD80/86＋ were significantly increased (30.1 vs. 49.9％ and 44.9 vs. 66.0％, respectively). Also, the difference in expressions of SWC1high SWC9low on BMHC-imDC from SWC1lowSWC9high on macrophages can be used as selective markers between these two antigen-presenting cells. Thus, we successfully established the in vitro differentiation method to generate large amounts of swine BMHC-imDC and BMHC-mDC, which can be used as screening cells for related immune studies.