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影响肾脏冷冻保存的因素。II. 三种冷冻保护剂与三种溶媒溶液联合应用于未冷冻兔皮质切片的毒性作用

Factors influencing renal cryopreservation. II. Toxic effects of three cryoprotectants in combination with three vehicle solutions in nonfrozen rabbit cortical slices.

作者信息

Clark P, Fahy G M, Karow A M

出版信息

Cryobiology. 1984 Jun;21(3):274-84. doi: 10.1016/0011-2240(84)90323-7.

Abstract

The [K+]/[Na+] ratio of rabbit renal cortical slices was used to examine, at 25 degrees C, the effects on viability of three cryoprotectant agents (CPA) (dimethyl sulfoxide (Me2SO), ethylene glycol, and glycerol) in combination with three vehicle solutions (Krebs-Henseleit (K-H), solution A, and RPS-2). Viability assessment by [K+]/[Na+] for all test solutions was made after incubating the slices in modified Cross-Taggart solution (C-T). With K-H and solution A, all concentrations of ethylene glycol and glycerol resulted in lowered ratios, whereas with Me2SO, concentrations greater than 1.4 M are required to reduce [K+]/[Na+]. With RPS-2 no decrease in the ratios was found until concentrations greater than 2.8 M were reached for all three CPAs. Binding of Me2SO to albumin, studied using [14C]Me2SO, was inhibited by RPS-2 when compared to K-H. Introduction and removal of Me2SO at 10 degrees C allowed an improvement in viability, at higher Me2SO concentrations, as compared to 25 degrees C.

摘要

在25℃下,利用兔肾皮质切片的[K⁺]/[Na⁺]比值来研究三种冷冻保护剂(CPA)(二甲基亚砜(Me₂SO)、乙二醇和甘油)与三种溶剂溶液(克氏-亨氏(K-H)溶液、A溶液和RPS-2)联合使用时对活力的影响。在改良的克罗斯-塔格特溶液(C-T)中孵育切片后,通过[K⁺]/[Na⁺]对所有测试溶液进行活力评估。对于K-H溶液和A溶液,所有浓度的乙二醇和甘油都会导致比值降低,而对于Me₂SO,需要大于1.4 M的浓度才能降低[K⁺]/[Na⁺]。对于RPS-2,直到三种CPA的浓度都达到大于2.8 M时,才发现比值降低。与K-H溶液相比,用[¹⁴C]Me₂SO研究发现,RPS-2可抑制Me₂SO与白蛋白的结合。与25℃相比,在10℃下引入和去除Me₂SO可在较高Me₂SO浓度下提高活力。

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