Mechanical interactions between ice crystals and red blood cells during directional solidification.
Experiments in which red blood cells were frozen on a directional solidification stage under a microscope show that there is a mechanical interaction between ice crystals and cells in which cells are pushed and deformed by the ice crystals. The mechanical interaction occurs during freezing of cells in physiological saline and is significantly inhibited by the addition of 20% v/v glycerol to the solution. The addition of osmotically insignificant quantities of antifreeze proteins from the winter flounder or ocean pout to the physiological saline with 20% v/v glycerol generates strong mechanical interactions between the ice and the cells. The cells were destroyed during freezing in physiological saline, survived freezing in physiological saline with glycerol, and were completely destroyed by the addition of antifreeze proteins to the solution with glycerol. The difference in cell survival through freezing and thawing appears to be related, in part, to the habit of ice crystal growing in the suspension of red blood cells and the nature of mechanical interaction between the ice crystal and the cells. This suggests that mechanical damage may be a factor during cryopreservation of cells.
Ishiguro
H
H
Institute of Engineering Mechanics, University of Tsukuba, Japan.
Rubinsky
B
B
D000818:Animals; D021301:Antifreeze Proteins; D001696:Biomechanical Phenomena; D001793:Blood Preservation / Q000009:adverse effects / Q000379:methods*; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D003460:Crystallization; D004912:Erythrocytes / Q000187:drug effects / Q000648:ultrastructure; D005069:Evaluation Studies as Topic; D005432:Flounder; D005990:Glycerol; D006023:Glycoproteins / Q000494:pharmacology; D006461:Hemolysis; D006801:Humans; D007053:Ice; D066298:In Vitro Techniques; D012965:Sodium Chloride
D021301:Antifreeze Proteins; D003451:Cryoprotective Agents; D006023:Glycoproteins; D007053:Ice; D012965:Sodium Chloride; D005990:Glycerol