Document Detail

Laser-assisted cryopreservation of single human spermatozoa in cell-free zona pellucida.
MedLine Citation:
PMID:  9949889     Owner:  NLM     Status:  MEDLINE    
Improved procedure for efficient cryopreservation of single human spermatozoa in cell-free human zona pellucida is reported. We used a diode laser system for efficient and precise creation of a single hole into the zona pellucida of a degenerated or immature human oocyte. This allowed the extraction of the cytoplasm using a micropipette with a diameter of 10-15 microns. Through the same opening, human spermatozoa were inserted into the empty zona. We used motile and laser immobilized spermatozoa. Immobilized sperm were obtained by a single laser irradiation delivered in the vicinity of the sperm tail prior to insertion. This new immobilization procedure was shown to have no deleterious effect on membrane integrity and sperm viability. Following sperm transfer into the zona, the laser-drilled hole was closed with an oil droplet which was expelled from the micropipette during withdrawal to avoid loss of spermatozoa. This facilitated detection of the otherwise translucent zona during the cryopreservation procedure. After thawing, all cryopreserved zonae (20/20) could be successfully retrieved. Spermatozoa were recovered from the zona pellucida through the hole used for insertion. The rate of sperm recovery for initially motile spermatozoa was 80% vs. 92% for laser immobilized spermatozoa. Sperm viability was 81% and 84%, respectively, detected by a Hoechst stain. This technique makes cryopreservation of single human spermatozoa easy and feasible and appears beneficial for couples with severe male infertility and for those facing repeated surgical sperm extraction.
M Montag; K Rink; U Dieckmann; G Delacrétaz; H van der Ven
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Andrologia     Volume:  31     ISSN:  0303-4569     ISO Abbreviation:  Andrologia     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-03-16     Completed Date:  1999-03-16     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0423506     Medline TA:  Andrologia     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  49-53     Citation Subset:  IM    
Department of Endocrinology and Reproductive Medicine, University of Bonn, Germany.
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MeSH Terms
Semen Preservation*
Zona Pellucida*

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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