Freezing human platelets with 6 percent dimethyl sulfoxide with removal of the supernatant solution before freezing and storage at-80°C without postthaw processing
Freezing human platelets with 6 percent dimethyl sulfoxide with removal of the supernatant solution before freezing and storage at-80°C without postthaw processing
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DOI：
10.1111/j.1537-2995.2005.00647.x
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发表时间：
2005-12-01
期刊：
影响因子：
2.9
通讯作者：
Khuri, S
中科院分区：
文献类型：
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作者：
Valeri, CR;Ragno, G;Khuri, S
背景：血小板 (PLT) 可以用 6% 二甲基亚砜 (DMSO) 在 -80 摄氏度下冷冻长达 2 年。该方法已通过浓缩 PLT 并在冷冻前去除上清液进行了修改。 研究设计和方法：将在 22 摄氏度下保存最多 5 天的高产去白细胞 PLT 分为三个等体积：一个在 -80 摄氏度下用 6% DMSO 冷冻，解冻、洗涤并重悬于血浆中（旧方法使用 DMSO）；另一个用 6% DMSO 在 -80 摄氏度下冷冻，解冻、洗涤并重悬于血浆中（旧方法使用 DMSO）。第二次用6% DMSO处理，浓缩除去上清液DMSO，-80℃冷冻，解冻，用0.9% NaCl稀释（DMSO新方法）；第三个用不含 DMSO 的 0.9% NaCl 处理，浓缩除去上清液，-80°C 冷冻，解冻，用 0.9% NaCl 稀释（不含 DMSO 的新方法）。结果：使用 5% PLT 微粒，用 DMSO 旧方法冷冻的 PLT 冻融洗涤回收率为 74 +/- 2%。 7% PLT 微粒（使用 DMSO 的新方法）的冻融回收率为 94 +/- 2%，15% PLT 微粒（不含 DMSO 的新方法）的冻融回收率为 69 +/- 9%。洗涤后的 PLT 中的总 DMSO 为 400 mg，冷冻前浓缩的 PLT 中的 DMSO 为 600 mg。采用DMSO新方法冷冻并输注到正常志愿者体内的PLT体内回收率为30%，寿命为7天。结论：冷冻前浓缩PLT无需解冻后洗涤，简化了程序。通过这种方法冷冻的 PLT 比通过旧方法冷冻的 PLT 具有更多的 GPIb 减少和膜联蛋白 V 结合增加的 PLT。
BACKGROUND: Platelets (PLTs) can be frozen with 6 percent dimethyl sulfoxide (DMSO) at -80 degrees C for up to 2 years. This method has been modified by concentrating the PLTs and removing the supernatant before freezing.STUDY DESIGN AND METHODS: High-yield leukoreduced PLTs stored at 22 degrees C for up to 5 days were divided into three equal volumes: one was frozen with 6 percent DMSO at -80 degrees C, thawed, washed, and resuspended in plasma (old method with DMSO); the second was treated with 6 percent DMSO, concentrated to remove the supernatant DMSO, frozen at -80 degrees C, thawed, and diluted with 0.9 percent NaCl (new method with DMSO); and the third was treated with 0.9 percent NaCl without DMSO, concentrated to remove the supernatant solution, frozen at -80 degrees C, thawed, and diluted with 0.9 percent NaCl (new method without DMSO).RESULTS: Freeze-thaw-wash recovery of PLTs frozen by the old method with DMSO was 74 +/- 2 percent with 5 percent PLT microparticles. Freeze-thaw recovery was 94 +/- 2 percent with 7 percent PLT microparticles (new method with DMSO) and 69 +/- 9 percent with 15 percent PLT microparticles (new method without DMSO). Total DMSO in washed PLTs was 400 and 600 mg in PLTs concentrated before freezing. In vivo recovery of PLTs frozen by the new method with DMSO and transfused into normal volunteers was 30 percent and the life span was 7 days.CONCLUSION: Concentrating PLTs before freezing simplified the procedure by eliminating postthaw washing. PLTs frozen by this method had more PLTs with reduced GPIb and increased annexin V binding than those frozen by the old method.