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Titolo:
Comparisons of oocyte maturation times and of three methods of sperm preparation for their effects on the production of goat embryos in vitro
Autore:
Rho, GJ; Hahnel, AC; Betteridge, KJ;
Indirizzi:
Univ Guelph, Ontario Vet Coll, Dept Biomed Sci, Guelph, ON N1G 2W1, CanadaUniv Guelph Guelph ON Canada N1G 2W1 omed Sci, Guelph, ON N1G 2W1, Canada Gyeongsang Natl Univ, Coll Vet Med, Chinju, South Korea Gyeongsang Natl Univ Chinju South Korea ll Vet Med, Chinju, South Korea
Titolo Testata:
THERIOGENOLOGY
fascicolo: 3, volume: 56, anno: 2001,
pagine: 503 - 516
SICI:
0093-691X(20010801)56:3<503:COOMTA>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
PERCOLL-GRADIENT CENTRIFUGATION; BOVINE FOLLICULAR OOCYTES; IN-VITRO; INVITRO FERTILIZATION; SWIM-UP; MEIOTIC COMPETENCE; HUMAN-SPERMATOZOA; SEMEN PREPARATION; TERM DEVELOPMENT; CULTURE;
Keywords:
goat; oocyte; IVM-IVF; sperm treatment; Percoll density-gradient;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Betteridge, KJ Univ Guelph, Ontario Vet Coll, Dept Biomed Sci, Guelph, ON N1G 2W1, Canada Univ Guelph Guelph ON Canada N1G 2W1 h, ON N1G 2W1, Canada
Citazione:
G.J. Rho et al., "Comparisons of oocyte maturation times and of three methods of sperm preparation for their effects on the production of goat embryos in vitro", THERIOGENOL, 56(3), 2001, pp. 503-516

Abstract

Various times of in vitro maturation of oocytes, and three methods of separating spermatozoa. from frozen-thawed semen (Percoll density-gradient centrifugation, swim-up, and glass-wool filtration), were compared for their effects on goat embryo production in vitro. Cumulus-oocyte-complexes (COCs) from abattoir ovaries were matured in M199 supplemented with 10% fetal calf serum and hormones. In Experiment 1, COCs were fixed at 4 h intervals from 0 to 27 h of culture to assess oocyte nuclear maturation. A higher proportion cultured for 27 h than for 24 h were in Metaphase II (27/37, 73% vs. 18/33, 55%; P < 0.05). In Experiment 2, the effects of separation methods on total numbers and numbers of membrane-intact spermatozoa, and the acrosome reaction were compared. Total numbers after Percoll density-gradient centrifugation were <similar to>4 times higher than after swim-up and similar to2 times higher than after glass-wool filtration (P < 0.001). Progression of the acrosome reaction was not affected differentially. In Experiments 3 and 4, after 27 h of culture the COCs were inseminated with sperm isolated by the three methods. In Experiment 3, presumptive zygotes were examined for pronucleus (PN) formation at 6, 12, 18 and 24 h post-insemination. At 12 h, male PN formation rate from Percoll-treated spermatozoa was higher than fromsperm subjected to swim-up and glass-wool treatments (20/37,54% vs. 6/37, 16% and 6/38, 16%, respectively; P < 0.001). In Experiment 4, embryos were compared for cleavage at 48 h and development into blastocysts, hatching rates and cell number at 192 h. The rates of cleavage and blastocyst formation in the Percoll-treated group were higher (P < 0.05) than in the swim-up and glass-wool groups (62% and 18% vs. 50% and 11%, and 45% and 9%, respectively). Similarly, the mean cell number in the Percoll group was higher (P <0.05) than in the swim-up and glass-wool groups (167 +/- 5 vs. 149 +/- 4 and 126 +/- 4, respectively). We conclude that Percoll density-gradient centrifugation is superior to the other two methods for separating goat spermatozoa from frozen-thawed semen in preparation for IVF. (C) 2001 by Elsevier Science Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/11/20 alle ore 15:21:05