Document Detail

Comparison of permeating and nonpermeating cryoprotectants for mouse sperm cryopreservation.
MedLine Citation:
PMID:  11336487     Owner:  NLM     Status:  MEDLINE    
Mouse sperm has proven to be more difficult to cryopreserve than sperm of other mammalian species. Published reports show that only three cryoprotectant agents (CPAs), alone or combined, have been studied: glycerol and dimethyl sulfoxide (DMSO), as permeating agents, and raffinose, as a nonpermeating agent. To date, the most consistent results for mouse sperm cryopreservation have been achieved by use of raffinose/skim milk as cryoprotectant with rapid cooling at 20 degrees C per minute. In this study, we compared the cryoprotection provided by permeating (glycerol, formamide, propanediol, DMSO, adonitol) or nonpermeating (lactose, raffinose, sucrose, trehalose, d-mannitol) compounds for freezing mouse sperm. Different solutions were made using 3% skim milk solution as the buffer or extender in which all different cryoprotectant agents were dissolved at a concentration of 0.3 M, with a final osmolality of approx. 400 mOsm. Sperm samples from CB6F1 (hybrid) and C57BL/6J (inbred) mice collected directly into each CPA were frozen/thawed under identical conditions. After thawing and CPA elimination (centrifugation) raffinose (59%), trehalose (61%), and sucrose (61%) sustained the best motility (P = < 0.1) of the nonpermeating agents, whereas the best of the permeating agents was DMSO (42%). Membrane integrity was analyzed and showed that the simple exposure (prefreeze) to sugars was less harmful than the exposure to glycols. Coincidentally, sperm frozen in trehalose (41%), raffinose (40.5%), and sucrose (37.5%) were the samples less injured among all different postthawed CPA tested. The in vitro fertilization results demonstrated that hybrid mouse spermatozoa frozen with sugars (lactose 80%, raffinose 80%, trehalose 79% of two-cell embryos production) were more fertile than those frozen with glycols (glycerol 11%).
J M Sztein; K Noble; J S Farley; L E Mobraaten
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Publication Detail:
Type:  Comparative Study; In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cryobiology     Volume:  42     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2001 Feb 
Date Detail:
Created Date:  2001-05-04     Completed Date:  2001-07-19     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  28-39     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA.
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MeSH Terms
Cell Survival / drug effects
Cryopreservation / methods*
Cryoprotective Agents / pharmacology
Embryo Transfer
Fertilization in Vitro
Mice, Inbred C57BL
Semen Preservation / methods*
Sperm Motility / drug effects
Spermatozoa / drug effects
Grant Support
Reg. No./Substance:
0/Cryoprotective Agents; 0/Solutions

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