利用过期血小板浓缩物生产人血小板裂解液作为T47D细胞增殖培养基

Diani Mentari, Relita Pebrina, Diah Nurpratami
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引用次数: 2

摘要

背景:浓缩血小板(PC)的保质期很短(5天)。过期PC不能用于临床目的。PC用于人血小板裂解物(HPL)的生产,其在增加T47D细胞增殖方面比FBS更有效。未见使用过期PC生产HPL的报道。本研究旨在探讨使用过期PC(储存时间>5天)生产的HPL是否可以促进体外T47D细胞的增殖。材料和方法:使用保质期为7天和11天的过期PC样品,通过冻融法生产HPL。测定pH、总蛋白含量、葡萄糖和白蛋白水平。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑(MTT)法测定hpl处理的T47D细胞的增殖率和倍增时间。结果:HPL生产后,葡萄糖水平受pH的影响(p=0.003),白蛋白水平受总蛋白含量的影响(p=0.030)。HPL保存7 d和11 d,细胞增殖率分别是10%胎牛血清的1.41倍和1.80倍。过期PC产生的HPL未引起细胞形态异常。在本研究中,葡萄糖水平影响细胞增殖(p=0.030)。高葡萄糖水平抑制T47D细胞增殖。结论:过期PC可提高细胞增殖速度,缩短细胞倍增时间,可作为生产HPL的潜在材料。关键词:细胞增殖,人血小板裂解液,浓缩血小板,血小板,T47D
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Utilization of Expired Platelet Concentrate for Production of Human Platelet Lysate as a Medium for T47D Cell Propagation
Background: Platelet concentrate (PC) has a short shelf life (5 days). Expired PC cannot be used for clinical purposes. PC is used for human platelet lysate (HPL) production, which was found to be more effective than FBS at increasing T47D cell proliferation. HPL production using expired PC has not been reported. This study aimed to investigate whether the use of HPL produced from expired PC (storage duration >5 days) can increase the proliferation of T47D cells in vitro.Materials and methods: Expired PC samples with a shelf life of 7 and 11 days were used to produce HPL via freeze/thaw method. pH, total protein content, glucose and albumin levels were measured. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure proliferation rate and doubling time of HPL-treated T47D cells.Results: After HPL production, the glucose level was influenced by the pH (p=0.003), and albumin level was influenced by total protein content (p=0.030). HPL stored for 7 and 11 days increased cell proliferation rate by 1.41 and 1.80 times higher than 10% FBS, respectively. HPL produced from expired PC did not cause morphological abnormality of the cells. In this study, the glucose levels affected cell proliferation (p=0.030). High glucose levels inhibited T47D cell proliferation.Conclusion: Expired PC can be used as a potential material for HPL production, since HPL produced from expired PC increases cell proliferation rate and shortens cell doubling time.Keywords: cell proliferation, human platelet lysate, platelet concentrate, thrombocyte, T47D
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