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目的:建立一种简单快速的体外扩增人外周血γδΤ细胞的方法,以便进一步对γδΤ细胞生物学特性和肿瘤免疫治疗的研究。方法:取健康人外周血100mL,分离单个核细胞,置于含100mL/L小牛血清,50mL/L人AB血清,异戊烯焦磷酸(2μg/L)和IL-2(100IU/mL)RPMI1640培养液中培养,进行细胞毒杀伤活性测定和流式细胞术分析。结果:细胞培养10d时γδΤ细胞从扩增前4·21%增加到70·35%,增殖倍数可达80倍,悬浮生长γδΤ细胞CD44表达为86·39%、贴壁生长γδΤ细胞为94·0%。效靶细胞比为40:1时,对K562细胞和SGC-7901胃腺癌细胞的杀伤活性分别为75%和61%。结论:用一种简单、快速和特异性的体外扩增人外周血中γδΤ细胞新方法,培养的γδΤ细胞具有较强的抗肿瘤活性。
OBJECTIVE: To establish a simple and rapid method for the expansion of human peripheral blood γδT cells in vitro, in order to further study the biological characteristics of γδT cells and tumor immunotherapy. Methods: Mononuclear cells (PBMCs) were isolated from 100 mL of peripheral blood of healthy volunteers and cultured in 100 mL / L fetal bovine serum, 50 mL / L human AB serum, 2 g / L isopentenyl pyrophosphate and 100 IU / RPMI1640 culture medium for cytotoxic activity assay and flow cytometry analysis. Results: The number of γδT cells increased from 4.21% to 70.35% at pre-expansion stage, and the multiplication rate was up to 80 times. The expression of CD44 in suspension-grown γδT cells was 86.39% 0%. When the target cell ratio was 40: 1, the killing activity against K562 cells and SGC-7901 gastric adenocarcinoma cells was 75% and 61%, respectively. CONCLUSION: The γδT cells cultured in vitro have a strong anti-tumor activity using a simple, rapid and specific method to amplify γδT cells in human peripheral blood in vitro.