Pegg D E, Hunt C J, Fong L P
MRC Medical Cryobiology Group, University Department of Surgery, Cambridge.
Cell Biophys. 1987 Apr;10(2):169-89. doi: 10.1007/BF02797398.
The process of cryopreservation subjects cells to gross changes in the composition of the solution that surrounds them, changes that cause the cells first to shrink and then to swell by an osmotic mechanism. Empirical methods have been developed that permit many cells to survive freezing and thawing, but the cornea, which is crucially dependent upon the function of its endothelial monolayer, has proved quite refractory. In this paper we explore the osmotic response of the corneal endothelium of the rabbit to solutions ranging in osmolality from 0.25 to 8.6 X isotonic. Boyle van't Hoff behavior was observed between 0.43 and 8.6 X isotonic, and there was an apparent nonosmotic volume of 33.6%. However, ultrastructural damage was observed at the limits of this range, and it appeared that the tolerated range was 0.64-4.4 X isotonic. We show the extent to which dimethyl sulfoxide (Me2SO) would be expected to moderate changes in volume during freezing and suggest that its initial concentration should be at least 2M to prevent excessive shrinkage. We also show that cell swelling during removal of Me2SO is especially likely to be hazardous.
冷冻保存过程会使细胞所处的周围溶液成分发生显著变化，这些变化会导致细胞先通过渗透机制收缩，然后膨胀。目前已经开发出一些经验方法，可使许多细胞在冻融后存活，但对于严重依赖其内皮单层功能的角膜来说，这些方法却难以奏效。在本文中，我们研究了兔角膜内皮对渗透压范围在0.25至8.6倍等渗的溶液的渗透反应。在0.43至8.6倍等渗之间观察到了玻意耳-范特霍夫行为，并且存在33.6%的表观非渗透体积。然而，在该范围的极限处观察到了超微结构损伤，似乎可耐受范围是0.64至4.4倍等渗。我们展示了二甲基亚砜（Me2SO）在冷冻过程中预期可缓解体积变化的程度，并表明其初始浓度应至少为2M，以防止过度收缩。我们还表明，在去除Me2SO期间细胞肿胀尤其可能具有危险性。
