De Loecker R, Goossens W, Van Duppen V, Verwilghen R, De Loecker W
Afdelingen Hematologie en Biochemie, Faculteit Geneeskunde, Katholieke Universiteit te Leuven, Belgium.
Cryobiology. 1993 Jun;30(3):279-85. doi: 10.1006/cryo.1993.1026.
Red blood cells frozen in 1.7 M and particularly in 2.2 M of glycerol retain a high degree of integrity upon thawing as long as the dilution procedure of the cryoprotectant is slow and preferentially compensated by the addition of sorbitol. As the nonpenetrating cryoprotectant sorbitol induces initial cell shrinkage, cell swelling upon dilution of the cryoprotectants may not lead to hemolysis. However, rapid dilution of glycerol even with buffer containing up to 0.50 M of sorbitol cannot be achieved without provoking considerable hemolysis. Due to the relative slow rate at which glycerol leaves the cells, membrane damage to the younger cell populations remains considerable and is even more pronounced in the older cell groups. The dramatic osmotic changes occurring during the dilution process lead to the formation of aberrant cell populations as demonstrated by the red cell size frequency distribution curves.
冷冻于1.7M尤其是2.2M甘油中的红细胞，只要冷冻保护剂的稀释过程缓慢且优先通过添加山梨醇来补偿，解冻时就能保持高度的完整性。由于非渗透性冷冻保护剂山梨醇会引起细胞初始收缩，冷冻保护剂稀释时的细胞肿胀可能不会导致溶血。然而，即使使用含有高达0.50M山梨醇的缓冲液，快速稀释甘油也会引发相当程度的溶血。由于甘油离开细胞的速度相对较慢，较年轻细胞群体的膜损伤仍然相当严重，在较老的细胞群体中甚至更明显。稀释过程中发生的剧烈渗透变化导致异常细胞群体的形成，红细胞大小频率分布曲线证明了这一点。
