Yan Xi1, Tianyu Miao2, Like Wan1, Yuanhu Wang1, Ting Feng2, Tianxiang Gong1, Mingyuan Li3. 1. Chengdu Blood Center, Sichuan University, Chengdu 610041, China. 2. West China School of Medicine, Sichuan University, Sichuan University, Chengdu 610041, China. 3. West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Abstract
OBJECTIVE: To compare the efficiency of pamidronate (PAM) and isopentenyl pyrophosphate (IPP) to stimulate γδ T cell expansion from human peripheral blood and explore the optimized expansion conditions. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Paque gradient centrifugation, and then cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, IPP (1.0, 5.0, 10.0, 15.0 μg/mL) or PAM (2.0, 5.0, 8.0, 12.0 μg/mL), and IL-2 (100.0, 200.0, 500.0 IU/mL). The cells were observed and collected. The number and proportion of CD3⁺TCRδ2⁺ γδ T cells stimulated by PAM or IPP in total lymphocytes were evaluated by flow cytometry and the expansion efficiency was calculated. RESULTS: After 14 days, the ratios of γδ T cells in total lymphocytes in IPP group and PAM group increased to 81.3% and 78.5%, respectively. This indicated that both IPP and PAM could effectively stimulate γδ T cell expansion and there was no significant difference in the efficiency of expansion between the two groups (P>0.05). CONCLUSION: PAM has the similar ability with IPP to stimulate γδ T cell expansion in vitro. PAM could become more economical and practical choice for stimulating γδ T cell expansion.
OBJECTIVE: To compare the efficiency of pamidronate (PAM) and isopentenyl pyrophosphate (IPP) to stimulate γδ T cell expansion from human peripheral blood and explore the optimized expansion conditions. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Paque gradient centrifugation, and then cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, IPP (1.0, 5.0, 10.0, 15.0 μg/mL) or PAM (2.0, 5.0, 8.0, 12.0 μg/mL), and IL-2 (100.0, 200.0, 500.0 IU/mL). The cells were observed and collected. The number and proportion of CD3⁺TCRδ2⁺ γδ T cells stimulated by PAM or IPP in total lymphocytes were evaluated by flow cytometry and the expansion efficiency was calculated. RESULTS: After 14 days, the ratios of γδ T cells in total lymphocytes in IPP group and PAM group increased to 81.3% and 78.5%, respectively. This indicated that both IPP and PAM could effectively stimulate γδ T cell expansion and there was no significant difference in the efficiency of expansion between the two groups (P>0.05). CONCLUSION:PAM has the similar ability with IPP to stimulate γδ T cell expansion in vitro. PAM could become more economical and practical choice for stimulating γδ T cell expansion.