Document Detail


Aseptic technology of vitrification of human pronuclear oocytes using open-pulled straws.
MedLine Citation:
PMID:  15528262     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The aim of this study was to compare the viability of human pronuclear oocytes subjected to vitrification using cooling by direct submerging of open-pulled straws in liquid nitrogen versus vitrification by cooling of open-pulled straws located inside a closed 0.5 ml straw (aseptic system). METHODS: Two- and three-pronuclei stage oocytes (n=114) were cryopreserved in super-open-pulled straws by vitrification in 20% ethylene glycol +20% dimethylsulphoxide (DMSO) + osmotic active and neutral non-permeable cryoprotectants with a four-step exposure in 20, 33, 50 and 100% vitrification solution for 2, 1 and 1 min, and 30-50 s, respectively at room temperature, and plunging into liquid nitrogen. Oocytes of group 1 (n=42) were rapidly cooled at a speed of 20,000 degrees C/min by direct plunging of open-pulled straws into liquid nitrogen. Oocytes of group 2 (n=44) were first located in 0.5 ml straws, which were closed at both sides by metal balls, and then plunged into liquid nitrogen. This method resulted in a cooling speed of 200 degrees C/min. For both groups, oocytes were thawed rapidly at a speed of 20 000 degrees C/min using an identical protocol. Oocytes subsequently were expelled into a graded series of sucrose solutions (1.0, 0.75, 0.5, 0.25 and 0.12 mol/l) at 2.5 min intervals. RESULTS: Oocyte development up to expanded blastocyst stage after in vitro culture was 15% in group 1, 14% in group 2 and 29% in an untreated control group. CONCLUSION: The deposition of human pronuclear oocytes in open-pulled straws which are placed inside a hermetically closed container guarantees a complete isolation of oocytes from liquid nitrogen and avoids potential contamination by pathogenic microorganisms. The combination of direct plunging of this container into liquid nitrogen and rapid warming makes this process as efficient as conventional vitrification.
Authors:
V Isachenko; M Montag; E Isachenko; V Zaeva; I Krivokharchenko; R Shafei; H van der Ven
Publication Detail:
Type:  Journal Article     Date:  2004-11-04
Journal Detail:
Title:  Human reproduction (Oxford, England)     Volume:  20     ISSN:  0268-1161     ISO Abbreviation:  Hum. Reprod.     Publication Date:  2005 Feb 
Date Detail:
Created Date:  2005-01-25     Completed Date:  2005-06-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8701199     Medline TA:  Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  492-6     Citation Subset:  IM    
Affiliation:
Department of Gynaecological Endocrinology and Reproductive Medicine, University of Bonn, Bonn, Germany. vovaisachenko@yahoo.com
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MeSH Terms
Descriptor/Qualifier:
Asepsis / instrumentation*,  methods*
Cell Nucleus
Cell Survival
Cryopreservation / instrumentation*,  methods*
Female
Fertilization in Vitro
Humans
Nitrogen
Oocytes / cytology*
Chemical
Reg. No./Substance:
7727-37-9/Nitrogen

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


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