体外培养人单个核细胞分泌IFN-γ水平的观察

Study on lever of IFN-γ secreted by human mononuclear cells cultured in vitro

目的对体外培养的人脐带血和外周血单个核细胞(MNC)以植物血凝素(PHA)刺激,观察培养上清中IFNγ含量变化。方法常规Ficoll离心法分离人MNC,接种24孔培养板,同时加入PHA,体外培养5d,计数不同培养时间的细胞总数;ELISA法测定不同培养时间上清中的IFNγ含量。结果经PHA刺激后,脐血MNC数量约扩增了12倍,单位细胞量所分泌的IFNγ是健康人血浆水平的30倍。而健康人外周血MNC数量约扩增了5倍。单位细胞量所分泌的IFNγ是健康人血浆水平的15倍。结论与人外周血MNC相比,人脐带血MNC对PHA刺激具有良好的反应性,经PHA刺激后可分泌更高水平的IFNγ,表明其在移植物抗白血病(GVL)效应中起一定作用。

Objective To observe the change of IFN-γ secreted from mononuclear cells (MNC) in human cord blood (UCB) and peripheral blood (PB) by stimulation of phytohemagglutinin (PHA) in vitro.Method MNC from UCB and PB were stimulated by PHA and cultured for 5 days in vitro.The cells and culture supernatants were collected.The number of cells was calculated.ELISA measured the level of IFN-γ in the culture supernatants.Result The number of MNC in UCB increased by 12-fold.The level of IFN-γ secreted by MNC in UCB was 30-fold higher than that in plasma of health subjects.The number of PB MNC increased by 5-fold.The level of IFN-γ of PB MNC was 15-fold higher than that in plasma of health subjects.Conclusion The mononuclear cells in human UCB were sensitive to PHA in vitro.This culture method for MNC can be used in anti-tumor immunotherapy.