Effects of Percoll separation, cryoprotective agents, and temperature on plasma membrane permeability characteristics of murine spermatozoa and their relevance to cryopreservation

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 ch...

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Bibliographic Details
Published in:Biology of reproduction 1999-10, Vol.61 (4), p.1031-1041
Main Authors: Phelps, M.J, Liu, J, Benson, J.D, Willoughby, C.E, Gilmore, J.A, Critser, J.K
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Membrane Permeability - drug effects
Cell Separation - methods
Cryopreservation
Cryoprotective Agents - pharmacology
density gradient centrifugation
dimethyl sulfoxide
ethylene glycol
Fundamental and applied biological sciences. Psychology
glycerol
hydraulic conductivity
Male
Mammalian male genital system
mathematical models
membrane permeability
Mice
Models, Biological
Morphology. Physiology
motility
plasma membrane
Povidone
propylene glycol
Silicon Dioxide
simulation models
Sperm Motility
spermatozoa
Spermatozoa - cytology
strain differences
Temperature
Vertebrates: reproduction
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Summary: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 (L p ) of murine spermatozoa. Both Percoll separation and cryoprotective agents (CPAs) decreased the L p . However, these effects were not additive. Experiment 3 was performed to determine the effect of temperature on L p in the presence of cryoprotectants (L p CPA ), cryoprotectant permeability (P CPA ), and the reflection coefficient (σ) 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.
ISSN:0006-3363
1529-7268
DOI:10.1095/biolreprod61.4.1031