Analysis of “solution effects” injury: Rabbit renal cortex frozen in the presence of dimethyl sulfoxide |
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Authors: | Gregory M. Fahy |
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Affiliation: | American National Red Cross, Blood Services Laboratories, Cryobiology Laboratory, 9312 Old Georgetown Rd., Bethesda, Maryland 20014 U.S.A. |
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Abstract: | Slices of rabbit renal cortex were frozen in 0.64 or 1.92 M dimethyl sulfoxide (Me2SO) to various subzero temperatures, thawed, and assayed for viability. Salt and Me2SO concentrations were calculated and correlated with the injury taking place during freezing. In separate experiments, slices were treated with NaCl or Me2SO in concentrations sufficient to simulate the exposure brought about as a result of freezing. The effects of these treatments on cortical viability were compared with the results of freezing to equivalent concentrations of either NaCl or Me2SO. The results show that whereas slices will tolerate exposure to at least six times the isotonic concentration of NaCl at 0 °C, they are unable to tolerate even three times the isotonic salt concentration when frozen in 1.92 M Me2SO. They can, however, tolerate 3 × NaCl when frozen in 0.64 M Me2SO. Freezing damage did not depend upon the amount of ice formed per se, since slices frozen in the low concentration of Me2SO tolerated removal of about 75% of the initial fluid content of the system, whereas slices frozen in 1.92 M Me2SO did not tolerate an identical removal of unfrozen solution. It was found that treatment of slices with high concentrations of Me2SO at subzero temperatures in accordance with Elford's application (14) of Farrant's method (20) produced damage which correlated approximately with the damage observed when the same concentrations of Me2SO were produced by freezing. It is concluded that most of the damage caused by freezing in 1.92 M Me2SO is produced either directly or indirectly by Me2SO. Possible mechanisms for this injury are discussed. |
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