冻干保护液用于较大容量冻干血小板生长因子活性的保护及其对人静脉内皮细胞增殖的作用

The effects of a protective agent on platelet growth factor activity and proliferation of HUVECs in large volume freeze-dried medium

目的研制用于较大容量(50 mL)冻干血小板生长因子的冻干保护液,探讨其对人静脉内皮细胞(HUVECs)增殖的作用。方法采集12位健康志愿者全血,并使用血液成分分离机手工分离血小板。将血小板汇集后,调整血小板计数约至1000×10^9/L。将调整好计数的血小板血浆分为5份,每份50 mL。分组:A组为新鲜血小板组;B组为冻干保护液处理组;C组为无冻干保护液处理组;D组为冻干保护液处理保存3个月组;E组为无冻干保护液保存3个月组。ELISA法检测血小板中生长因子的含量,CCK8法检测HUVECs增殖率。结果生长因子含量与细胞增殖结果一致,C组(TGF:192216.680±109705.640,VEGF:78.157±8.831,PDGF:16985.43±1031.256,bFGF:178.969±13.282,CCK-8:1.008±0.151),D组(TGF:246626.621±17039.413,VEGF:85.694±12.292,PDGF:15472.204±378.222,bFGF:116.736±16.149,CCK-8:0.681±0.035),E组(TGF:150862.825±9023.450,VEGF:46.945±1.311,PDGF:9389.033±262.193,bFGF:98.691±9.679,CCK-8:0.037±0.029)与A组(TGF:425458.163±25862.627,VEGF:383507.356±50170.545,PDGF:26531.360±1188.531,bFGF:178.969±13.282,CCK-8:1.258±0.047)相比,均显著降低(P<0.05);C、D、E与B组(TGF:383507.356±50170.545,VEGF:119.250±10.928,PDGF:23850.094±2185.510,bFGF:172.993±23.169,CCK-8:1.237±0.045)相比,均显著降低(P<0.05)E组与D组相比,E组显著降低(P<0.05)。结论研制的冻干保护液对较大容量冻干血小板中生长因子活性及其促进HUVECs生长有较好的保护作用。

Objective To investigate the effects of freeze-dried protective agent prepared by our group on the activities of four main growth factors in large volume(50 mL)freeze-dried platelets and their effects on the proliferation of HUVECs.Methods Whole blood was collected from 12 healthy volunteers.Platelet was separated by blood component separator,and adjusted to about 1000×10^9 L.The adjusted plasma of rich platelet was divided into 5 parts(50 mL each):group A:fresh platelet group;group B:treated with lyophilized protective solution;Group C:treated with no freeze-dried protective liquid;Group D:preserved with freeze-dried protective liquid for three months;group E:preserved without freeze-dried protective liquid for three months.The content of growth factor in platelets was detected by ELISA and the proliferation rate of HUVECs was detected by CCK-8 method.Results Compared with group A(TGF:425458.163±25862.627,VEGF:383507.356±50170.545,PDGF:26531.360±1188.531,bFGF:178.969±13.282,and CCK-8:1.258±0.047),group C(TGF:192216.680±109705.640,VEGF:78.157±8.831,PDGF:16985.43±1031.256,bFGF:178.969±13.282,and CCK-8:1.008±0.151),group D(TGF:246626.621±17039.413,VEGF:85.694±12.292,PDGF:15472.204±378.222,bFGF:116.736±16.149,and CCK-8:0.681±0.035),group E(TGF:150862.825±9023.450,VEGF:46.945±1.311,PDGF:9389.033±262.193,bFGF:98.691±9.679,and CCK-8:0.037±0.029)were significantly lower(P<0.05).Compared with Group B(TGF:383507.356±50170.545,VEGF:119.250±10.928,PDGF:23850.094±2185.510,bFGF:172.993±23.169,and CCK-8:1.237±0.045),the other freeze-dried group were significantly decreased(P<0.05).Compared with group D,group E was significantly decreased(P<0.05).Conclusion The freeze-dried protective agent prepared by our research group has a good protective effect on the growth factor activity and promotes HUVECs proliferation in large volume freeze-dried platelets.