^(60)Co照射法饲养细胞的制备

Preparation of Feeder Cells by Irradiation with ^(60)Co

目的 :选择合适的60 钴 (60 Co)照射剂量制备饲养细胞。方法 :采用 0Gy、2Gy、5Gy、1 0Gy、2 0Gy、30Gy的60钴 (60 Co)照射人的外周血单个核细胞 (PBMC) ,然后将其继续培养 ,加入CD3 单克隆抗体 (OKT3 )、白细胞介素 - 2(IL - 2 )、植物血球凝集素 (PHA)等 ,观察照射后第 1天、第 7天及第 1 5天细胞的凋亡情况及活细胞的细胞周期分布情况。结果 :γ射线照射剂量的加大会导致PBMC增殖能力的下降、存活细胞 (Annexin -V-/PI-)的减少、凋亡 (Annexin -V+ /PT-)与死亡 (Annexin -V+ /PT+ )的细胞数增加。结论 :采用60 Co照射法制备饲养细胞时 ,1

Objective To select an appropriate dosage of irradiation with 60 Co for preparation of feeder cells. Methods Human peripheral blood monocyte (PBMC) irradiated with 60 Co at the dosage of 0Gy、2Gy、5Gy、10Gy、20Gy and 30Gy respectively were cultured in virto and stimulated with OKT 3、 IL-2 and PHA. Flow cytometric analyses of apoptosis rate and cell cycle of irradiated PBMC were performed on D1、D7 and D15. Results Increasing the irradiation dosage would lead to more inhibition of proliferation of PBMC, with decreasing number of living cells (Annexin-V -/PI -) and increasing number of apoptoitic (Annexin-V +/PI -) and dead (Annexin-V +/PI +) cells up to 10Gy. Further increasing would make little difference. Conclusion For preparationof feeder cells from PBMC, irradiation with 60 Co at the dosage of 10Gy may be most appropriate.