Document Detail


Recovery, survival and functional evaluation by transplantation of frozen-thawed mouse germ cells.
MedLine Citation:
PMID:  14990537     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Establishing a successful method for testicular stem cell transplantation of frozen-thawed testicular cells would be of immense benefit to boys with childhood cancer undergoing a sterilizing treatment. In this study, we evaluated different cryopreservation protocols in a mouse model by means of testicular germ cell transplantation (TGCT), in order to establish an optimal freezing protocol. METHODS AND RESULTS: In a first series of experiments, we compared an uncontrolled protocol with 1.5 mol/l dimethyl sulphoxide (DMSO) versus a controlled long protocol (cooling to -80 degrees C) and observed a better viability with the latter protocol (36% versus 48%, P < 0.05). We then compared survival after two thawing methods (37 degrees C water versus ice water) in either a DMSO- or an ethylene glycol (EG)-based protocol, and found no difference. In order to evaluate the functional capacity of the cryopreserved testicular suspension, TGCT was performed with both fresh and frozen-thawed suspensions. In 90% of the successfully injected testes, spermatogenesis was reinitiated using fresh suspensions. In contrast, this figure was only 12.5 and 22.7% after cryopreservation, for the short controlled EG protocol and the uncontrolled DMSO protocol, respectively. CONCLUSION: Reinitiation of spermatogenesis is possible after cryopreservation of testicular germ cell suspensions. Although cell survival was acceptable, our results after TGCT show that our protocols need further improvement.
Authors:
V Frederickx; A Michiels; E Goossens; G De Block; A C Van Steirteghem; H Tournaye
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2004-02-27
Journal Detail:
Title:  Human reproduction (Oxford, England)     Volume:  19     ISSN:  0268-1161     ISO Abbreviation:  Hum. Reprod.     Publication Date:  2004 Apr 
Date Detail:
Created Date:  2004-03-22     Completed Date:  2004-11-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8701199     Medline TA:  Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  948-53     Citation Subset:  IM    
Affiliation:
Centre for Reproductive Medicine and Research Laboratories for Reproductive Medicine, University Hospital and Medical School, Vrije Universiteit Brussel, Laarbeeklaan 101, B-1090 Brussels, Belgium. veerle.frederickx@az.vub.ac.be
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Survival / drug effects
Cryopreservation*
Cryoprotective Agents / pharmacology
Cryptorchidism / genetics
Dimethyl Sulfoxide / pharmacology
Ethylene Glycol / pharmacology
Heterozygote
Male
Mice
Mice, Transgenic
Semen Preservation*
Spermatozoa / drug effects,  physiology*,  transplantation*
Time Factors
Tissue Donors
Chemical
Reg. No./Substance:
0/Cryoprotective Agents; 107-21-1/Ethylene Glycol; 67-68-5/Dimethyl Sulfoxide

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