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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 observe the changes of the levels of tumor necrosis factor-α (TNF-α) in human peripheral blood and cord blood mononuclear cells (MNCs) and the culture supernatant stimulated by lipopolysaccharide (LPS). Methods: Human peripheral blood and umbilical cord blood MNC were isolated by routine Ficoll centrifugation and seeded into 24-well plates. LPS was added into the culture medium for 5 days. The total number of cells in different culture time was counted. The expression of TNF- α content. Results: After LPS stimulation, the number of cord blood MNC expanded about 12 times, and the amount of TNF-α secreted per unit cell volume was 30 times higher than that of healthy people. The number of MNC in peripheral blood of healthy people increased about 5 times, The amount of TNF-α secreted by the cells is 15 times higher than that of healthy people. Conclusion: Compared with human peripheral blood MNC, human cord blood MNC has a better response to LPS stimulation and a higher level of TNF-α secretion, providing a new idea and approach for the clinical application of TNF-α in the treatment of malignant tumors.