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Registro Completo |
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Biblioteca(s): |
Embrapa Caprinos e Ovinos. |
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Data corrente: |
17/07/2013 |
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Data da última atualização: |
08/09/2021 |
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Tipo da produção científica: |
Resumo em Anais de Congresso |
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Autoria: |
BRACKETT, B. G.; KESKINTEPE, L.; SIMPLÍCIO, A. A.; LUVONI, G. G. |
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Afiliação: |
Department of Physiology & Pharmacology, College of Veterinary Medicine, UGA, Athens, GA 30602, USA; Department of Physiology & Pharmacology, College of Veterinary Medicine, UGA, Athens, GA 30602, USA; AURINO ALVES SIMPLÍCIO, CNPC; Instituto di Clinical Ostetrica e Ginecologica Veterinaria, Universita? degli Studi di Milano, Milano, Italy. |
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Título: |
Influences of culture components on the development of bovine blastocysts in defined conditions. |
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Ano de publicação: |
1997 |
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Fonte/Imprenta: |
Theriogenology, v. 47, n. 1, p. 274, Jan., 1997. |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract: Experiments were conducted to determine influences on in vitro production (IVP) of bovine embryos of glucose (glc), citrate (c), glutathione (GSH), and growth factors, EGF and PDGF, during 9 d culture in defined conditions. Oocytes, aspirated from follicles (2-5 mm) at slaughter, with homogeneous cytoplasm and 2-3 layers of cumulus cells were washed twice in Tyrodes with 400 mug PVA/ml and 112 mug sodium pyruvate/ml and incubated in 100 mul of mTCM 199 + 10 mug oFSH and 5 mug bLH (NHPP, NIDDK, NICHD, USDA)/ml for 24 h before insemination. Then, after 6 h co-culture with heparin-capacitated sperm, ova were cultured in 50 mul drops of either c-SOF+NEA (control) (Biol.Reprod. 52:1410-1417) or c-SOF+NEA without glc and c (I), or without glc (II) for 5 d; then, developing embryos were transferred into c-SOF+NEA +/- GSH for 4 d. In another trial EGF or PDGF were added to c-SOF+NEA for the last 4 d. Culture media were renewed (50% by volume) every 24 h. Proportions of oocytes that cleaved (C) by 48 h, reached morulae (M) by 120 h, blastocysts (B) by 168 h, and expanded B (EB) by 216 h, and, of M that proceeded to B and EB were analyzed by ''Sigma Stat'' by means of ANOVA; differences among the treatments were analyzed by Bonferroni-t test. Removal of either glc and c, or glc, compromised results, e.g. C of 72.3 % for I, 64.8 % for II, and 86.1 % for control. Addition of GSH 5 d after beginning embryo culture was not beneficial for B development after initial culture in I, 31.3%, or II, 25.2%. Significantly more B and EB were obtained after 9 d in c-SOF+NEA (40.8 % and 34.1 %, P<0.05). Evaluation of development rates of d 5 M to B stages on d 7, 8, 9 revealed more M cultured in I followed by no GSH reached growing B stage by d 7 (27.3 %) than for other experimental groups (11.6 % for I with GSH, 14.9 % for II without GSH, and 18.9 % for II with GSH), but development was faster in c-SOF+NEA (39.6 %) (P<0.05). Early inclusion of citrate was found to enhance M to B development from d 5 to d 8 and, d 9 regardless of other treatments after d 5. In another experiment, different concentrations (0.05, 0.5, 5.0 ng/ml) of EGF or PDGF were added to c-SOF+NEA for the last 4 d of culture. The lowest concentration of either EGF or PDGF resulted in lower (P<0.05) blastocyst development (33.3 % for EGF and 46.6 % for PDGF) compared to the highest concentration (60.0 % and 80.0 %, respectively). EB development was significantly higher (P<0.05) in media containing either growth factor at 5 ng/ml; 32 of 60 M (53.3 %) reached EB with EGF, and 28 (46.6 %) of 60 M reached EB with PDGF present during the final 4 d. Results demonstrated positive influences of glc and c, absence of beneficial effect of GSH, variable patterns in M to B progression, and efficacy of EGF and PDGF in enhancing EB development. MenosAbstract: Experiments were conducted to determine influences on in vitro production (IVP) of bovine embryos of glucose (glc), citrate (c), glutathione (GSH), and growth factors, EGF and PDGF, during 9 d culture in defined conditions. Oocytes, aspirated from follicles (2-5 mm) at slaughter, with homogeneous cytoplasm and 2-3 layers of cumulus cells were washed twice in Tyrodes with 400 mug PVA/ml and 112 mug sodium pyruvate/ml and incubated in 100 mul of mTCM 199 + 10 mug oFSH and 5 mug bLH (NHPP, NIDDK, NICHD, USDA)/ml for 24 h before insemination. Then, after 6 h co-culture with heparin-capacitated sperm, ova were cultured in 50 mul drops of either c-SOF+NEA (control) (Biol.Reprod. 52:1410-1417) or c-SOF+NEA without glc and c (I), or without glc (II) for 5 d; then, developing embryos were transferred into c-SOF+NEA +/- GSH for 4 d. In another trial EGF or PDGF were added to c-SOF+NEA for the last 4 d. Culture media were renewed (50% by volume) every 24 h. Proportions of oocytes that cleaved (C) by 48 h, reached morulae (M) by 120 h, blastocysts (B) by 168 h, and expanded B (EB) by 216 h, and, of M that proceeded to B and EB were analyzed by ''Sigma Stat'' by means of ANOVA; differences among the treatments were analyzed by Bonferroni-t test. Removal of either glc and c, or glc, compromised results, e.g. C of 72.3 % for I, 64.8 % for II, and 86.1 % for control. Addition of GSH 5 d after beginning embryo culture was not beneficial for B development after initial culture in I,... Mostrar Tudo |
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Palavras-Chave: |
Tecnologia de embrião. |
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Thesagro: |
Bovino; Embrião animal. |
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Thesaurus Nal: |
Cattle; Embryo culture; embryo transfer. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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Marc: |
LEADER 03493nam a2200217 a 4500
001 1962286
005 2021-09-08
008 1997 bl uuuu u00u1 u #d
100 1 $aBRACKETT, B. G.
245 $aInfluences of culture components on the development of bovine blastocysts in defined conditions.$h[electronic resource]
260 $aTheriogenology, v. 47, n. 1, p. 274, Jan., 1997.$c1997
520 $aAbstract: Experiments were conducted to determine influences on in vitro production (IVP) of bovine embryos of glucose (glc), citrate (c), glutathione (GSH), and growth factors, EGF and PDGF, during 9 d culture in defined conditions. Oocytes, aspirated from follicles (2-5 mm) at slaughter, with homogeneous cytoplasm and 2-3 layers of cumulus cells were washed twice in Tyrodes with 400 mug PVA/ml and 112 mug sodium pyruvate/ml and incubated in 100 mul of mTCM 199 + 10 mug oFSH and 5 mug bLH (NHPP, NIDDK, NICHD, USDA)/ml for 24 h before insemination. Then, after 6 h co-culture with heparin-capacitated sperm, ova were cultured in 50 mul drops of either c-SOF+NEA (control) (Biol.Reprod. 52:1410-1417) or c-SOF+NEA without glc and c (I), or without glc (II) for 5 d; then, developing embryos were transferred into c-SOF+NEA +/- GSH for 4 d. In another trial EGF or PDGF were added to c-SOF+NEA for the last 4 d. Culture media were renewed (50% by volume) every 24 h. Proportions of oocytes that cleaved (C) by 48 h, reached morulae (M) by 120 h, blastocysts (B) by 168 h, and expanded B (EB) by 216 h, and, of M that proceeded to B and EB were analyzed by ''Sigma Stat'' by means of ANOVA; differences among the treatments were analyzed by Bonferroni-t test. Removal of either glc and c, or glc, compromised results, e.g. C of 72.3 % for I, 64.8 % for II, and 86.1 % for control. Addition of GSH 5 d after beginning embryo culture was not beneficial for B development after initial culture in I, 31.3%, or II, 25.2%. Significantly more B and EB were obtained after 9 d in c-SOF+NEA (40.8 % and 34.1 %, P<0.05). Evaluation of development rates of d 5 M to B stages on d 7, 8, 9 revealed more M cultured in I followed by no GSH reached growing B stage by d 7 (27.3 %) than for other experimental groups (11.6 % for I with GSH, 14.9 % for II without GSH, and 18.9 % for II with GSH), but development was faster in c-SOF+NEA (39.6 %) (P<0.05). Early inclusion of citrate was found to enhance M to B development from d 5 to d 8 and, d 9 regardless of other treatments after d 5. In another experiment, different concentrations (0.05, 0.5, 5.0 ng/ml) of EGF or PDGF were added to c-SOF+NEA for the last 4 d of culture. The lowest concentration of either EGF or PDGF resulted in lower (P<0.05) blastocyst development (33.3 % for EGF and 46.6 % for PDGF) compared to the highest concentration (60.0 % and 80.0 %, respectively). EB development was significantly higher (P<0.05) in media containing either growth factor at 5 ng/ml; 32 of 60 M (53.3 %) reached EB with EGF, and 28 (46.6 %) of 60 M reached EB with PDGF present during the final 4 d. Results demonstrated positive influences of glc and c, absence of beneficial effect of GSH, variable patterns in M to B progression, and efficacy of EGF and PDGF in enhancing EB development.
650 $aCattle
650 $aEmbryo culture
650 $aembryo transfer
650 $aBovino
650 $aEmbrião animal
653 $aTecnologia de embrião
700 1 $aKESKINTEPE, L.
700 1 $aSIMPLÍCIO, A. A.
700 1 $aLUVONI, G. G.
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Registro original: |
Embrapa Caprinos e Ovinos (CNPC) |
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