摘要
目的研究血清浓度和培养基对人γδT细胞增殖、表型和杀伤活性的影响。方法采用AIMV培养基,添加不同浓度胎牛血清对γδT细胞进行培养,比较细胞增殖、表型和杀伤活性。在此基础上,采用不同培养基(AIMV、Op Tmizer)对γδT细胞进行培养,比较细胞增殖、表型和杀伤活性。自动细胞计数仪检测γδT细胞数;流式细胞术(FCM)检测γδT细胞表型;CCK-8试剂盒检测γδT细胞对胃癌SGC-7901的杀伤效应。结果采用AIMV培养基,无血清条件下细胞无法生长,添加5%或10%胎牛血清(FBS)培养,细胞增殖优于添加1%FBS(P=0.024和P=0.006)或10%NBS(P=0.043和P=0.006),具有统计学意义,杀伤活性不具有统计学差异;采用Op Tmizer培养基,无血清及添加1%FBS培养均优于添加5%FBS的AIMV培养基,其中,1%FBS与AIMV5%FBS组相比具有显著性差异(P=0.030)。结论 Op Tmizer培养基比AIMV培养基更适用于临床过继免疫治疗用γδT细胞的无血清培养。
Objective The aim of the study is to investigate the roles of two media with varied concentrations of bovine serum in biological characteristics of the γδT cells, including the proliferation, phenotypes and the cytolytic activity. Methods The AIMV medium containing varied concentrations of FBS was applied to culture the γδT cells respectively and the bio-functional characteristics of the cells like the proliferation, phenotypes, and cytolytic activity were identified. Based on the preliminary findings, we applied two media(AIMV, Op Tmizer) to generate the γδT cells respectively and examined the proliferation, phenotypes, and cytolytic activity. The number of the γδT cells was counted by automated cell counter; the phenotype-related surface molecules were determined by FCM; the cytolytic activity to kill SGC-7901 cells was assayed by CCK-8 kit. Results Cultured in AIMV medium, γδT cells were unable to survive in serum-dependent manner, in which case, the cells grew far better in the medium with 5% FBS or 10% FBS than those in the medium with 1% FBS or 10% NBS. In contrast, the expansion of γδT cells in Op Tmizer medium with 1% FBS or serum-free is superior to that of those in AIMV medium with 5% FBS. Conclusion Our findings suggest Op Tmizer medium in the γδT cell ex-vivo expansion may be superior to AIMV medium.
出处
《中华临床医师杂志（电子版）》
CAS
2015年第23期101-104,共4页
Chinese Journal of Clinicians(Electronic Edition)
关键词
血清
培养基
细胞增殖
表型
γδT细胞
杀伤活性
Serum
Culture media
Cell proliferation
Phenotype
γδT cells
Cytolytic activity
