Radojcić L, Vukotić-Maletić V, Balint B
Klinicki centar, Beograd, Vojnomedicinska akademiji, Beograd Ginekolosko odeljenje.
Med Pregl. 1998 Jan-Feb;51(1-2):29-36.
The aim of various cryopreservation procedures is to minimize cell injuries during the freeze-thaw cycle (cryoinjuries). Generally, the cell damage during freezing and thawing procedures may be the results of: (a) extensive cellular dehydration (solution effect) and/or (b) intracellular ice crystallization/recrystallization (mechanical cell damage). Two independent mechanisms are involved. They can act simultaneously, leading to cytolysis. The first one is expressed primarily during low rate freezing, and the second one during rapid freezing. Thus, determination and use of the optimal cooling velocity, specific for each type of isolated cells, should be considered. Finally, a higher degree of cell destruction has been documented when the transition period from liquid to solid phase (release of the fusion heat) is prolonged. CRYOPROTECTIVE AGENTS: For successful cell cryopreservation, cryoprotectants are needed. They decrease the osmotic gradient and the vapor pressure difference between the intra- and extracellular area. Adequate choice of the most suitable type and concentration of cryoprotective agent is important for the required cell recovery after thawing. There are several well known protocols for obtaining cryopreservation of isolated cells using different cryoprotectants. Glycerol, dimethyl sulfoxide (DMSO) and propanediol sucrose are commonly used as cryoprotectants, though in different concentrations. Glycerol, a trihydric alcohol, is a clear, colorless fluid. Pharmacologically, it is relatively inert. DMSO is a colorless liquid with a sulphur-like smell and has several medical uses. It is highly polar and dissolves many water- and lipid-soluble substances. DMSO given intravenously may cause nausea, vomiting, local vasospasm and an objectionable garlic-like odor and taste. HUMAN SPERM, OVA AND EMBRYOS CRYOPRESERVATION: Despite the fact that cryopreservation procedures of spermatozoa, ova and embryos are already in routine clinical use, some questions related to the optimal cooling velocity during controlled-rate freezing and the choice of the most effective, either penetrating (glycerol, dimethyl sulfoxide) and/or non-penetrating (hydroxyethyl starch) cryoprotective agent at the appropriate concentration are not resolved.
各种冷冻保存程序的目的是尽量减少冻融循环期间的细胞损伤（冷冻损伤）。一般来说，冷冻和解冻过程中的细胞损伤可能是以下结果：(a) 广泛的细胞脱水（溶液效应）和/或 (b) 细胞内冰晶形成/再结晶（机械性细胞损伤）。涉及两种独立的机制。它们可以同时起作用，导致细胞溶解。第一种主要在慢速冷冻过程中表现出来，第二种在快速冷冻过程中表现出来。因此，应考虑确定并使用针对每种分离细胞类型的最佳冷却速度。最后，当从液相到固相的过渡期（熔化热释放）延长时，已证明细胞破坏程度更高。
为了成功进行细胞冷冻保存，需要冷冻保护剂。它们可降低细胞内和细胞外区域之间的渗透梯度和蒸气压差。适当选择最合适的冷冻保护剂类型和浓度对于解冻后所需的细胞复苏很重要。有几种使用不同冷冻保护剂进行分离细胞冷冻保存的知名方案。甘油、二甲基亚砜 (DMSO) 和丙二醇蔗糖通常用作冷冻保护剂，不过浓度不同。甘油是一种三元醇，是一种清澈、无色的液体。在药理学上，它相对惰性。DMSO 是一种无色液体，有类似硫磺的气味，有多种医学用途。它具有高度极性，能溶解许多水溶性和脂溶性物质。静脉注射 DMSO 可能会引起恶心、呕吐、局部血管痉挛以及令人反感的大蒜样气味和味道。
人类精子、卵子和胚胎冷冻保存：尽管精子、卵子和胚胎的冷冻保存程序已在临床常规使用，但与控制速率冷冻期间的最佳冷却速度以及选择适当浓度的最有效穿透性（甘油、二甲基亚砜）和/或非穿透性（羟乙基淀粉）冷冻保护剂相关的一些问题尚未得到解决。
