Kheirolomoom Azadeh, Satpathy Gyana R, Török Zsolt, Banerjee Mitali, Bali Rachna, Novaes Roberta C, Little Erika, Manning Danielle M, Dwyre Denis M, Tablin Fern, Crowe John H, Tsvetkova Nelly M
Center for Biostabilization and Section of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA.
Cryobiology. 2005 Dec;51(3):290-305. doi: 10.1016/j.cryobiol.2005.08.003. Epub 2005 Sep 26.
In a previous report [Z. Török, G. Satpathy, M. Banerjee, R. Bali, E. Little, R. Novaes, H. Van Ly, D. Dwyre, A. Kheirolomoom, F. Tablin, J.H. Crowe, N.M. Tsvetkova, Preservation of trehalose loaded red blood cells by lyophilization, Cell Preservation Technol. 3 (2005) 96-111.], we presented a method for preserving human red blood cells (RBCs) by loading them with trehalose and then freeze-drying. We have now improved that method, based on the discovery that addition of phospholipid vesicles to the lyophilization buffer substantially reduces hemolysis of freeze-dried RBCs after rehydration. The surviving cells synthesize 2,3-DPG, have low levels of methemoglobin, and have preserved morphology. Among the lipid species we studied, unsaturated PCs were found to be most effective in suppressing hemoglobin leakage. RBC-vesicle interactions depend on vesicle size and structure; unilamellar liposomes with average diameter of less than 300 nm were more effective in reducing the hemolysis than multilamellar vesicles. Trehalose loaded RBCs demonstrated high survival and low levels of methemoglobin during 10 weeks of storage at 4 degrees C in the dry state when lyophilized in the presence of liposomes.
在之前的一份报告[Z. 托罗克、G. 萨帕西、M. 班纳吉、R. 巴利、E. 利特尔、R. 诺瓦斯、H. 范利、D. 德怀尔、A. 凯罗拉莫姆、F. 塔布林、J.H. 克罗、N.M. 茨韦特科娃，通过冻干保存负载海藻糖的红细胞，《细胞保存技术》3 (2005) 96 - 111。]中，我们提出了一种通过给人类红细胞（RBCs）负载海藻糖然后冻干来保存它们的方法。我们现在改进了该方法，这是基于以下发现：在冻干缓冲液中添加磷脂囊泡可显著减少复水后冻干红细胞的溶血。存活的细胞能合成2,3 - 二磷酸甘油酸（2,3 - DPG），高铁血红蛋白水平较低，且形态得以保留。在我们研究的脂质种类中，发现不饱和磷脂酰胆碱（PCs）在抑制血红蛋白泄漏方面最有效。红细胞与囊泡的相互作用取决于囊泡的大小和结构；平均直径小于300 nm的单层脂质体在减少溶血方面比多层囊泡更有效。当在脂质体存在的情况下冻干时，负载海藻糖的红细胞在4℃干燥状态下储存10周期间表现出高存活率和低水平的高铁血红蛋白。
