目的探讨外周血经3种不同抗凝剂处理后培养的自然杀伤(NK)细胞的增殖率和杀伤功能的影响研究。方法采集10例年龄(35±5)岁健康志愿者15 m L外周血,分别置于含乙二胺四乙酸二钾(EDTA-K2,EDTA组)、细胞保存液(细胞保存液组)和肝素钠(...目的探讨外周血经3种不同抗凝剂处理后培养的自然杀伤(NK)细胞的增殖率和杀伤功能的影响研究。方法采集10例年龄(35±5)岁健康志愿者15 m L外周血,分别置于含乙二胺四乙酸二钾(EDTA-K2,EDTA组)、细胞保存液(细胞保存液组)和肝素钠(肝素钠组)的抗凝管中。用淋巴细胞分离液收集单个核细胞,在NK细胞培养液中培养单个核细胞12 d,并观察细胞生长情况。在第4、6、8、10、12天时,收集细胞用CCK8法检测NK细胞的增殖情况。在细胞增殖最明显的第12天收集细胞,通过流式细胞仪检测各组NK细胞的颗粒酶、穿孔素和CD107a。台盼蓝染色检测细胞活性。通过CCK8法检测各组对肿瘤细胞株K562杀伤活性。结果通过细胞增殖发现,EDTA组NK细胞增殖不明显,肝素钠组和细胞保存液组增殖明显。第12天时检测发现,肝素钠组增殖倍数为128.15±6.38,细胞保存液组增殖倍数为107.51±5.70;肝素钠组优于细胞保存液组(t=6.20,P=0.00)。肝素钠组NK细胞纯度表达为(84.49±2.35)%,也明显高于细胞保存液组NK细胞表达[(77.63±1.37)%]。经台盼蓝染色检测,肝素钠组和细胞保存液组细胞活性率均为100%。通过流式细胞仪检测发现,肝素钠组NK细胞颗粒酶B、穿孔素和CD107a的表达都明显高于细胞保存液组(t=12.76、17.76、5.67,P=0.00、0.00、0.00)。对肿瘤细胞株K562杀伤结果显示,肝素钠组NK细胞杀伤活性(50.12%±3.42%)明显高于细胞保存液组(41.88%±2.47%)(t=4.78,P=0.001)。结论肝素钠和细胞保存液均可用于NK细胞培养的血液抗凝,但肝素钠组NK细胞的杀伤功能明显优于细胞保存液组。展开更多
肿瘤的发生发展与免疫系统关系密切。程序性死亡受体1(programmed death receptor-1,PD-1)主要表达于T细胞等免疫细胞表面。程序性死亡受体配体1(programmed cell death ligand-1,PD-L1)表达于多种肿瘤细胞表面,包括肺癌、黑色素瘤、卵...肿瘤的发生发展与免疫系统关系密切。程序性死亡受体1(programmed death receptor-1,PD-1)主要表达于T细胞等免疫细胞表面。程序性死亡受体配体1(programmed cell death ligand-1,PD-L1)表达于多种肿瘤细胞表面,包括肺癌、黑色素瘤、卵巢癌、肾癌、口腔鳞状细胞癌等,与PD-1结合后可抑制T细胞迁移、增殖和杀伤功能,实现肿瘤免疫逃逸。肿瘤细胞通过多种机制逃避免疫系统攻击,PD-1/PD-L1信号通路是其主要机制之一。展开更多
Objective To investigate the short- and long-term therapeutic efficacies of intravenous transplantation of bone marrow stem cells(MSCs) in rats with experimental myocardial infarction by metaanalysis.Methods Randomize...Objective To investigate the short- and long-term therapeutic efficacies of intravenous transplantation of bone marrow stem cells(MSCs) in rats with experimental myocardial infarction by metaanalysis.Methods Randomized controlled trials were systematically searched from Pub Med,Science Citation Index(SCI),Chinese journal full-text database(CJFD) up to December 2014.While the experimental groups(MSCs groups) were injected MSCs intravenously,the control groups were injected Delubecco's minimum essential medium(DMEM) or phosphate buffered saline(PBS).Subgroup analysis for each outcome measure was performed for the observing time point after the transplantation of MSCs.Weighted mean differences(WMD) and 95% confidence intervals(CI) were calculated for outcome parameters including ejection fraction(EF) and fractional shortening(FS),which were measured by echocardiogram after intravenous injection and analyzed by Rev Man 5.2 and STATA 12.0.Results Data from 9 studies(190 rats) were included in the meta-analysis.As compared to the control groups,the cardiac function of the experimental groups were not improved at day 7(EF:WMD=0.08,95%CI-1.32 to 1.16,P>0.01; FS:WMD=-0.12,95%CI-0.90 to 0.65,P>0.01) until at day 14 after MSCs' transplantation(EF:WMD=10.79,95%CI 9.16 to 12.42,P<0.01; FS:WMD=11.34,95%CI 10.44 to 12.23,P<0.01),and it lasted 4 weeks or more after transplantation of MSCs(EF:WMD=13.94,95%CI 12.24 to 15.64,P<0.01; FS:WMD=9.64,95%CI 7.98 to 11.31,P<0.01).Conclusion The therapeutic efficacies of MSCs in rats with myocardid infarction become increasing apparent as time advances since 2 weeks after injection.展开更多
文摘目的探讨外周血经3种不同抗凝剂处理后培养的自然杀伤(NK)细胞的增殖率和杀伤功能的影响研究。方法采集10例年龄(35±5)岁健康志愿者15 m L外周血,分别置于含乙二胺四乙酸二钾(EDTA-K2,EDTA组)、细胞保存液(细胞保存液组)和肝素钠(肝素钠组)的抗凝管中。用淋巴细胞分离液收集单个核细胞,在NK细胞培养液中培养单个核细胞12 d,并观察细胞生长情况。在第4、6、8、10、12天时,收集细胞用CCK8法检测NK细胞的增殖情况。在细胞增殖最明显的第12天收集细胞,通过流式细胞仪检测各组NK细胞的颗粒酶、穿孔素和CD107a。台盼蓝染色检测细胞活性。通过CCK8法检测各组对肿瘤细胞株K562杀伤活性。结果通过细胞增殖发现,EDTA组NK细胞增殖不明显,肝素钠组和细胞保存液组增殖明显。第12天时检测发现,肝素钠组增殖倍数为128.15±6.38,细胞保存液组增殖倍数为107.51±5.70;肝素钠组优于细胞保存液组(t=6.20,P=0.00)。肝素钠组NK细胞纯度表达为(84.49±2.35)%,也明显高于细胞保存液组NK细胞表达[(77.63±1.37)%]。经台盼蓝染色检测,肝素钠组和细胞保存液组细胞活性率均为100%。通过流式细胞仪检测发现,肝素钠组NK细胞颗粒酶B、穿孔素和CD107a的表达都明显高于细胞保存液组(t=12.76、17.76、5.67,P=0.00、0.00、0.00)。对肿瘤细胞株K562杀伤结果显示,肝素钠组NK细胞杀伤活性(50.12%±3.42%)明显高于细胞保存液组(41.88%±2.47%)(t=4.78,P=0.001)。结论肝素钠和细胞保存液均可用于NK细胞培养的血液抗凝,但肝素钠组NK细胞的杀伤功能明显优于细胞保存液组。
文摘肿瘤的发生发展与免疫系统关系密切。程序性死亡受体1(programmed death receptor-1,PD-1)主要表达于T细胞等免疫细胞表面。程序性死亡受体配体1(programmed cell death ligand-1,PD-L1)表达于多种肿瘤细胞表面,包括肺癌、黑色素瘤、卵巢癌、肾癌、口腔鳞状细胞癌等,与PD-1结合后可抑制T细胞迁移、增殖和杀伤功能,实现肿瘤免疫逃逸。肿瘤细胞通过多种机制逃避免疫系统攻击,PD-1/PD-L1信号通路是其主要机制之一。
基金Supported by the Youth Project of National Natural Science Foundation(81100078)the Key Project of Chinese Ministry of Education(211207)+1 种基金Guangzhou Pearl River science and technology new star project plan(2012J2200063)Project of Guangdong Science and Technology Department(S2011040001392)
文摘Objective To investigate the short- and long-term therapeutic efficacies of intravenous transplantation of bone marrow stem cells(MSCs) in rats with experimental myocardial infarction by metaanalysis.Methods Randomized controlled trials were systematically searched from Pub Med,Science Citation Index(SCI),Chinese journal full-text database(CJFD) up to December 2014.While the experimental groups(MSCs groups) were injected MSCs intravenously,the control groups were injected Delubecco's minimum essential medium(DMEM) or phosphate buffered saline(PBS).Subgroup analysis for each outcome measure was performed for the observing time point after the transplantation of MSCs.Weighted mean differences(WMD) and 95% confidence intervals(CI) were calculated for outcome parameters including ejection fraction(EF) and fractional shortening(FS),which were measured by echocardiogram after intravenous injection and analyzed by Rev Man 5.2 and STATA 12.0.Results Data from 9 studies(190 rats) were included in the meta-analysis.As compared to the control groups,the cardiac function of the experimental groups were not improved at day 7(EF:WMD=0.08,95%CI-1.32 to 1.16,P>0.01; FS:WMD=-0.12,95%CI-0.90 to 0.65,P>0.01) until at day 14 after MSCs' transplantation(EF:WMD=10.79,95%CI 9.16 to 12.42,P<0.01; FS:WMD=11.34,95%CI 10.44 to 12.23,P<0.01),and it lasted 4 weeks or more after transplantation of MSCs(EF:WMD=13.94,95%CI 12.24 to 15.64,P<0.01; FS:WMD=9.64,95%CI 7.98 to 11.31,P<0.01).Conclusion The therapeutic efficacies of MSCs in rats with myocardid infarction become increasing apparent as time advances since 2 weeks after injection.
