Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2002-9-25
pubmed:abstractText
Efficient and dependable mouse cryopreservation methods are urgently needed because the production of mice with transgenes and disrupted and mutant genes is now commonplace. Preservation of these unique genomes provides an essential safeguard for future research. Unfortunately, mouse spermatozoa appear more vulnerable to freezing than other species, e.g., bovine and human. In this study, we examined the efficiency of intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) in generating embryos from mouse spermatozoa frozen with 18% raffinose and 3% skim milk for cryoprotection. A comparison was made between the inbred strain C57BL/6J, commonly used in mutagenic and transgenic studies, and a hybrid strain B6D2F1 (C57BL/6J x DBA/2J). C57BL/6J spermatozoa are known to be more sensitive to freezing than B6D2F1. Fertilization of oocytes after IVF was significantly lower with C57BL/6J spermatozoa when compared with B6D2F1 spermatozoa for both fresh and frozen spermatozoa (fresh, 89 vs. 55%; frozen, 56 vs. 9%). Freezing also reduced the fertility of B6D2F1 spermatozoa (89 vs. 56%). Fertilization improved dramatically after ICSI with fresh and frozen C57BL/6J spermatozoa (90 and 85%) and also with frozen B6D2F1 spermatozoa (87%). The development of two-cell embryos to the blastocyst stage was lower for C57BL/6J than B6D2F1 (42-61% and 84-98%) in all treatments but similar for embryos within each strain. The normality of chromosomes from fresh and frozen spermatozoa was assessed in oocytes prior to first cleavage. The majority of oocytes had normal chromosomes after IVF (98-100%) and ICSI (87-95%), indicating that chromosomal abnormalities were not responsible for the poorer development in vitro of C57BL/6J embryos. In conclusion, our data show that ICSI is a more efficient and effective technique than IVF for generating embryos from frozen spermatozoa. More important, ICSI is especially valuable for strains where IVF with fresh spermatozoa produces few or no embryos.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-3363
pubmed:author
pubmed:issnType
Print
pubmed:volume
67
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1278-84
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:12297546-Animals, pubmed-meshheading:12297546-Blastocyst, pubmed-meshheading:12297546-Cell Count, pubmed-meshheading:12297546-Chromosome Aberrations, pubmed-meshheading:12297546-Cryopreservation, pubmed-meshheading:12297546-Cryoprotective Agents, pubmed-meshheading:12297546-Culture Techniques, pubmed-meshheading:12297546-Embryonic and Fetal Development, pubmed-meshheading:12297546-Epididymis, pubmed-meshheading:12297546-Fertilization in Vitro, pubmed-meshheading:12297546-Hybridization, Genetic, pubmed-meshheading:12297546-Male, pubmed-meshheading:12297546-Mice, pubmed-meshheading:12297546-Mice, Inbred C57BL, pubmed-meshheading:12297546-Mice, Inbred DBA, pubmed-meshheading:12297546-Semen Preservation, pubmed-meshheading:12297546-Sperm Injections, Intracytoplasmic, pubmed-meshheading:12297546-Spermatozoa
pubmed:year
2002
pubmed:articleTitle
Intracytoplasmic sperm injection is more efficient than in vitro fertilization for generating mouse embryos from cryopreserved spermatozoa.
pubmed:affiliation
Institute for Biogenesis Research, University of Hawaii Medical School, Honolulu 96822, USA. szczygie@hawaii.edu
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S.