两种来源的树突状细胞体外培养及免疫学特征比较

Immunity character of dendritic cells derived from two blood

目的对脐血(UCB)和外周血(PB)体外诱导培养成树突状细胞(DC),并进行免疫学特征的比较,探索更好的DC来源。方法UCB和正常成人PB各10例为UCB组和PB组,分离出单个核细胞后,分别加入粒细胞巨细胞集落刺激因子(GM-CSF),白介素4(IL-4)和肿瘤坏死因子(TNF-α)培养基中,培养12d,每天用光镜观察细胞形态,于第3、6d和10d,用流式细胞仪进行CD11c、CD80、CD86及HLA-DR表型分析,于第10d透射电镜和扫描电镜观察细胞形态,并在第12d与淋巴细胞混合培养,对比两种源性DC激发淋巴细胞增殖能力。结果两者均诱导出具有典型形态的DC,UCB组诱导的DC细胞数量明显多于外周血诱导的细胞数(P<0.01)。激发淋巴细胞增殖能力,两者无区别(P>0.05)。结论UCB和PB中的MNs均可在体外诱导为成熟DC,UCB诱导的DC细胞数高于PB,两者激发淋巴细胞增殖能力相同,故UCB有可能成为免疫治疗DC的丰富来源。

Objective：To investigate how to induce dendritic cells（DC） from umbilical cord blood（UCB） and peripheral blood（PB） and test it＇ s immunological characteristics. Methods： Ten UCB and 10 normal adult PB were collected. The mononuclear cells （MNCs） of two groups（ UCB group and PB group） were separated and cultured with granulocyte - macrophage colony - stimulating factor （ GM - CSF）, interleukin - 4 （ IL - 4 ） and tumor necrosis factor- α（TNF- α） for 12 days. The cell configuration was observed by optical microscope every day. CD11c, CDSO,CD86 and HLA - DR were tested by flow cytometry at 3d, 6d, and 10d separately. The cell configuration was observed by transmission electric microscope and scan electric microscope at 10d. The DCs were cultured with lym- phocyte for testing the ability to stimulate lymphocyte proliferation at 12d. Results： DCs with typical morphological characteristics were induced successfully in two groups. The yields of DCs from UCB group were higher than that from PB group（ P 〈0.01 ）. DCs in two groups were not different in stimulating lymphocyte proliferation（ P 〉 0.05）. Conclusion： The MNs in UCB and PB may be induced into mature DC. The yields of DCs from UCB are higher than that from PB. DCs from the two sources have same ability in stimulating lymphocyte proliferation. Therefore, UCB may be valuable origin for DC immunotherapy.