体外脂多糖刺激人单个核细胞分泌TNF-α水平的研究

Study of human mononuclearcell secreting TNF-?琢 stimulated by lipopolysaccharide in vitro

目的:观察脂多糖(LPS)刺激体外培养的人外周血和脐带血单个核细胞(MNC)及培养上清中肿瘤坏死因子α(TNF-α)的含量变化。方法:常规Ficoll离心法分离人外周血和脐带血MNC,接种于24孔培养板,加入LPS,体外培养5天,计数不同培养时间的细胞总数;ELISA法测定不同培养时间培养上清中TNF-α含量。结果:经LPS刺激后,脐带血MNC数量约扩增了l2倍,单位细胞量所分泌的TNF-α是健康人血浆水平的30倍;健康人外周血MNC数量约扩增了5倍,单位细胞量所分泌的TNF-α是健康人血浆水平的l5倍。结论:与人外周血MNC相比,人脐带血MNC对LPS刺激具有更好的反应性,可分泌更高水平的TNF-α,为临床应用TNF-α治疗恶性肿瘤提供新的思路和途径。

Objective：To study the content of tumor neceosis factor-alpha（TNF-α） in the culture supernatant of mononuelearcell that was isolated from human peripheral blood and cord blood and stimulated by lipopolysaccharide （LPS） in vitro. Methods ：Fieoll centrifugalization method was used to isolate the mononueleareell fiom human periperal blood and cord blood,there after the monomueleareell was inoculated in 24-bor tissue culture plate and added with LPS respectively,and eultured for 5 days in vitro. The amount of cell in the different euhure times was counted. ELISA method was used to measure the content of TNF-α in the different culture times. Results：After the stimulation of LPS, the quantity of mononuelearcell in human cord blood expanded 12 times.TNF-α secreted by the unit cell quantity was 30 times that of normal healthy plasma;the quantity of mononuelearcell in human peripheral blood expanded 5 times ,the TNF-α secreted by the unit cell quantity was 15 times that of normal healthy plasma. Conclusion： Compared with the mononuelearcell of human peripheral blood,the mononuelearcell of human cord blood has a higher reaction to LPS stimulation ,and can secrete more TNF-α.