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a Cryobiology Research Institute, Wells Research Center, Indiana University Medical School, Indianapolis,Indiana 46202
b Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette,Indiana 47907
Cryopreservation of murine spermatozoa would provide an efficient method for preserving important genotypes. However, to date such methods have resulted in low survivals with significant variability. To address this issue, a series of five experiments was performed to determine the cryobiological characteristics of murine spermatozoa. Experiments 1 and 2 investigated the effect of Percoll separation on the hydraulic conductivity (Lp) of murine spermatozoa. Both Percoll separation and cryoprotective agents (CPAs) decreased the Lp. However, these effects were not additive. Experiment 3 was performed to determine the effect of temperature on Lp in the presence of cryoprotectants (LpCPA), cryoprotectant permeability (PCPA), and the reflection coefficient (
) in spermatozoa from both ICR and B6C3F1 mice. Permeability parameters decreased as temperature decreased, and permeability characteristics differed between strains. In experiments 4 and 5, theoretical simulations for CPA addition and removal were developed and empirically tested. Strain-specific methods for CPA addition and removal based upon the fundamental cryobiological characteristics of murine spermatozoa resulted in higher survivals than current methods or procedures, which were used as controls.
2 Correspondence: John K. Critser, Cryobiology Research Institute, Cancer Research Building, Wells Research Center, Room 454, 1044 West Walnut Street, Indianapolis, IN 46202. FAX: 317 274 8679; jcritser{at}iupui.edu
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