恒河猴骨髓来源未成熟树突状细胞的培养及鉴定

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (9): 1243-1248.

恒河猴骨髓来源未成熟树突状细胞的培养及鉴定

支良¹,陈鹏²,李朝战¹,李昌亮¹,关兴¹,褚朋¹,阿永俊²

1. 枣庄市立医院
2. 昆明医科大学第二附属医院

收稿日期:2014-12-04 修回日期:2015-04-27 出版日期:2015-09-05 发布日期:2015-09-07
通讯作者: 阿永俊 E-mail:997991476@qq.com
基金资助:
国家自然科学基金( 30960378), 国家自然科学基金(81160069) 内设研究机构云南省卫生厅研究项目(2011WS0084)

Culture and identification of immature dendritic cells from rhesus monkey myeloid

Received:2014-12-04 Revised:2015-04-27 Online:2015-09-05 Published:2015-09-07

摘要/Abstract

摘要： 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6d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。结论此方法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。

关键词: 关键词：恒河猴, 未成熟树突状细胞, 免疫耐受

Abstract: Objective To approach 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 Screen rhesus monkey bone marrow cells using a magnetic activated cell sorting (MACS) system 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 isolated CD34+ hematopoietic stem cells to induce bone marrow-derived DCs. Morphological changes of cells were observed under an electron microscope, and cell surface molecules were detected using flow cytometry after cells were stained with DC surface marker CD1a and mature 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 established a method for culturing highly purified rhesus monkey bone marrow-derived immature DCs (imDC). Characterization of imDCs and functional studies will be carried out shortly.

Key words: rhesus monkeys, immature dendritic cells, immune tolerance

中图分类号:

中图分类号： Q813.1+1 文献标识码：A

支良陈鹏李朝战李昌亮关兴褚朋阿永俊. 恒河猴骨髓来源未成熟树突状细胞的培养及鉴定[J]. 基础医学与临床, 2015, 35(9): 1243-1248.

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