CAJ | 학술논문

目的：建立一种体外诱导恒河猴骨髓CD34＋细胞分化为树突状细胞（ DC）的方法，并对其细胞形态及表面标志进行鉴定。方法用免疫磁珠分选系统（ MACS）分选恒河猴骨髓细胞，收集高纯度的CD34＋造血干细胞；加入重组粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素4（IL-4）及肿瘤坏死因子-α（TNF-α），诱导其分化为骨髓来源的树突状细胞，并采用电镜观察细胞形态。用树突状细胞表面标志CD1a及成熟树突状细胞（ mDC）表面标志CD83流式抗体染色后分析。结果细胞因子诱导6 d后成功培养出表面呈绒毛状和树枝状突起的典型DC样细胞，流式细胞术分析结果显示92.35％的细胞为DC，27.61％的细胞为mDC。结论用此方法可在体外培养出高纯度且典型的恒河猴骨髓源性未成熟树突状细胞（ imDC），为进一步研究其生物学特性及功能奠定了基础。
목적：건립일충체외유도항하후골수CD34＋세포분화위수돌상세포（ DC）적방법，병대기세포형태급표면표지진행감정。방법용면역자주분선계통（ MACS）분선항하후골수세포，수집고순도적CD34＋조혈간세포；가입중조립세포-거서세포집락자격인자（GM-CSF）화백세포개소4（IL-4）급종류배사인자-α（TNF-α），유도기분화위골수래원적수돌상세포，병채용전경관찰세포형태。용수돌상세포표면표지CD1a급성숙수돌상세포（ mDC）표면표지CD83류식항체염색후분석。결과세포인자유도6 d후성공배양출표면정융모상화수지상돌기적전형DC양세포，류식세포술분석결과현시92.35％적세포위DC，27.61％적세포위mDC。결론용차방법가재체외배양출고순도차전형적항하후골수원성미성숙수돌상세포（ imDC），위진일보연구기생물학특성급공능전정료기출。
Objective To develop a method for culturing dendritic cells ( DC ) from rhesus monkey bone marrow CD34 +cells, and identify the cells by morphological observation and cell surface marker detection .Methods To screen rhesus monkey bone marrow cells using a magnetic activated cell sorting (MACS) system then to collect highly purified CD34+hematopoietic stem cells .Recombinant granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-4 ( IL-4) and tumor necrosis factor-α( TNF-α) were applied to isolate CD 34+hematopoietic stem cells to induce bone marrow-derived DCs .Morphological change of cells was observed under an electron microscope , and cell surface molecules were detected using flow cytometry after cells were stained with DC surface marker CD 1a and ma-ture DC surface marker CD83.Results Typical DCs with surface villous and dendritic protrusions were identified 6 days after cytokine treatment .92.35%of the cultured cells were DC and 27.61%of the cultured cells were mature DCs ( mDC) .Conclusions We have established a method for culturing highly purified rhesus monkey bone marrow -derived immature DCs (imDC), which may function as a technine plateform to support research in future .