肿瘤相关巨噬细胞模型建立的实验研究

doi:10.3969/j.issn.1673-8640.2014.09.007

检验医学 ›› 2014, Vol. 29 ›› Issue (9): 897-802.DOI: 10.3969/j.issn.1673-8640.2014.09.007

肿瘤相关巨噬细胞模型建立的实验研究

张国良,刘鹥雯,何怡青,杨翠霞,王文涓,杜艳,高锋

上海交通大学附属第六人民医院中心实验室,上海 200233

收稿日期:2014-04-09 出版日期:2014-09-30 发布日期:2014-09-25
通讯作者: 高锋,联系电话:021-64369181-58702
作者简介:张国良,男,1987年生,学士,主要从事肿瘤免疫学研究
基金资助:
国家自然科学基金资助项目(81071814、81172027、81272479);上海市自然科学基金资助项目(12ZR1447400)

The experimental study of establishing tumor-associated macrophage model

ZHANG Guoliang, LIU Yiwen, HE Yiqing, YANG Cuixia, WANG Wenjuan, DU Yan, GAO Feng

Center for Clinical Laboratory, Shanghai Jiaotong University Affiliated Sixth People's Hospital, Shanghai 200233, China

Received:2014-04-09 Online:2014-09-30 Published:2014-09-25

摘要/Abstract

摘要：

目的建立肿瘤相关巨噬细胞(TAM)的体外分化模型。方法分离健康人外周血单核细胞进行体外培养;经白细胞介素4(IL-4)、白细胞介素13(IL-13)刺激后,利用流式细胞术检测细胞表面标志物CD14、CD204、CD206的表达改变;利用酶联免疫吸附试验(ELISA)检测单核细胞白细胞介素10(IL-10)分泌量的改变;继而将刺激后的单核细胞与肿瘤细胞共培养,采用Hoechst染色观察其对肿瘤细胞的杀伤情况。结果经IL-4和IL-13刺激后,单核细胞表面CD14的表达率未发生明显变化,而CD204和CD206的表达显著增高,同时IL-10的分泌量也显著增高;与肿瘤细胞共培养结果显示,经IL-4/IL-13刺激的单核细胞对肿瘤细胞的杀伤能力显著下降。结论 IL-4/IL-13可在体外诱导单核细胞呈现TAM的表型和功能,适用于TAM分化模型的建立,该模型可应用于针对TAM的相关研究。

Abstract:

Objective To establish the differentiation model of tumor-associated macrophages (TAM) in vitro. Methods Monocytes were isolated from the peripheral blood of healthy subjects and cultured in vitro. After stimulation by interleukin 4(IL-4) and interleukin 13(IL-13), the expression changes of molecule markers on monocytes were determined by flow cytometry, including CD14, CD204 and CD206. The levels of interleukin 10 (IL-10) secreted by monocytes weredetermined by enzyme-linked iummunosorbent assay (ELISA). After coculturing with monocytes, the apoptosis of tumor cells was investigated by Hoechst staining. Results After stimulation by IL-4 and IL-13, monocytes exhibited a markedly altered phenotype with retained CD14, the increasing expressions of CD204 and CD206 and the increasing amount of secreted IL-10. Moreover, when coculturing with tumor cells, the cytotoxity of monocytes on tumor cells was inhibited by IL-4/IL-13. Conclusions IL-4/IL-13 could induce monocytes to exhibit TAM phenotype and function, indicating IL-4/IL-13 could be applied to the establishment of TAM differentiation model, which can be used inthe research of TAM.

Key words: Tumor-associated macrophage, Interleukin 4, Interleukin 13, Monocyte, M2 macrophage

中图分类号:

R446.61

张国良,刘鹥雯,何怡青,杨翠霞,王文涓,杜艳,高锋. 肿瘤相关巨噬细胞模型建立的实验研究[J]. 检验医学, 2014, 29(9): 897-802.

ZHANG Guoliang, LIU Yiwen, HE Yiqing, YANG Cuixia, WANG Wenjuan, DU Yan, GAO Feng. The experimental study of establishing tumor-associated macrophage model[J]. , 2014, 29(9): 897-802.

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肿瘤相关巨噬细胞模型建立的实验研究

The experimental study of establishing tumor-associated macrophage model

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