细胞因子体外诱导脐带血单个核细胞向成熟肥大细胞分化  

In vitro differentiation of human umbilical cord blood mononuclear cells into mature mast cells induced by cytokines

作　　者：陈惠芳[1] 何颖[1] 张兰珍[2] 刘兆宇[1] 邹泽红[1] 陶爱林[1] 

机构地区：[1]广州医科大学附属第二医院广东省过敏反应与免疫重点实验室,呼吸疾病国家重点实验室,广东广州510260 [2]广州医科大学附属第二医院产科,广东广州510260

出　　处：《细胞与分子免疫学杂志》2015年第9期1255-1258,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家科技重大专项重大课题(2014ZX08011-005);国家科技重大专项重点课题(2014ZX08011-005 B);国家自然科学基金(81373128)

摘　　要：目的分离脐带血单个核细胞并进行体外定向诱导培养,获得人源化、纯度高的成熟肥大细胞。方法用重组人干细胞生长因子(rh SCF)、重组人白细胞介素6(rh IL-6)定向诱导从脐带血中分离的单个核细胞,使其分化成为成熟的肥大细胞。收集不同时间段的细胞,进行甲苯胺蓝染色并用流式细胞术检测其膜上FcεRⅠ受体,以鉴定其成熟程度。成熟的肥大细胞经过敏原激发后,检测细胞上清中的组织胺及类胰蛋白酶。结果在加入rh SCF和rh IL-6诱导2周以后,肥大细胞即开始表达FcεRⅠ受体,诱导3周后达到高峰,同时细胞质内嗜碱性颗粒增多。诱导成熟后的细胞经过敏原激发后,能有效释放类胰蛋白酶和组织胺。结论脐带血单个核细胞可定向诱导成为成熟的肥大细胞,可用于过敏原性的体外评价试验。Objective To isolate and induce human umbilical cord blood mononuclear cells to differentiate into mature mast cells of high purity. Methods Human umbilical cord blood mononuclear cells were differentiated into mature mast cells by the treatment of recombinant human stem cell factor（ rhSCF） and recombinant human interleukin 6（ rhIL-6）. The cultured cells at different time points were stained with toluidine blue for the detection of anti-FcεRⅠ and the maturity of mast cells was detected by flow cytometry. After the mature mast cells were stimulated with allergen,the levels of histamine and tryptase in the supernatant were determined. Results The cells started to express FcεRⅠ receptor after 2-week treatment of rh SCF and rh IL-6. After 3 weeks,the amount of FcεRⅠ receptor reached its peak accompanied by increased intracellular basophilic granules. The mature mast cells released tryptase and histamine effectively after allergen challenge. Conclusion Human umbilical cord blood mononuclear cells can be differentiated into mature mast cells by the treatment of rhSCF and rhIL-6. The mature mast cells may be used for the study of allergenicity in vitro.

关 键 词：脐血单个核细胞 肥大细胞 组织胺 类胰蛋白酶 

分 类 号：R392.8[医药卫生—免疫学]