ABSTRACT
O objetivo desse estudo foi comparar dois diferentes sistemas de incubação (incubadora convencional CONV e minibancada MINI) sobre a produção in vitro de embriões (PIVE) de bovinos. Para tanto, complexos cumulusoócito (CCOs) foram submetidos a maturação in vitro e posterior fecundação e cultivo in vitro em ambos os sistemas. As estruturas foram avaliadas pós-maturação e pós-cultivo, para avaliação de maturação nuclear e quantidade de células/blastocisto, respectivamente. Não foram observadas diferenças estatísticas (p>0,05) nos diferentes parâmetros estudados. Concluímos que ambos os sistemas testados demonstraram ser eficientes para PIVE de bovinos.(AU)
The objective of this study was to compare two different incubation systems (conventional incubator - CONV and minibank - MINI) on the in vitro production of bovine embryos (PIVE). For this purpose, cumulus cytotoxic complexes (CCOs) were submitted to in vitro maturation and subsequent fertilization and in vitro culture in both systems. The structures were evaluated post-maturation and post-culture, for evaluation of nuclear maturation and amount of cells / blastocyst, respectively. No statistical differences (p>0.05) were observed in the different parameters studied. We conclude that both tested systems proved to be efficient for bovine PIVE.(AU)
Subject(s)
Animals , Cattle , Embryo, Mammalian/embryology , Embryonic Development , In Vitro Techniques/veterinary , In Vitro Techniques/methods , Embryo Culture Techniques/veterinary , Embryo Culture Techniques/methods , Incubators/veterinary , Reproductive Techniques, Assisted/veterinaryABSTRACT
O objetivo desse estudo foi comparar dois diferentes sistemas de incubação (incubadora convencional CONV e minibancada MINI) sobre a produção in vitro de embriões (PIVE) de bovinos. Para tanto, complexos cumulusoócito (CCOs) foram submetidos a maturação in vitro e posterior fecundação e cultivo in vitro em ambos os sistemas. As estruturas foram avaliadas pós-maturação e pós-cultivo, para avaliação de maturação nuclear e quantidade de células/blastocisto, respectivamente. Não foram observadas diferenças estatísticas (p>0,05) nos diferentes parâmetros estudados. Concluímos que ambos os sistemas testados demonstraram ser eficientes para PIVE de bovinos.
The objective of this study was to compare two different incubation systems (conventional incubator - CONV and minibank - MINI) on the in vitro production of bovine embryos (PIVE). For this purpose, cumulus cytotoxic complexes (CCOs) were submitted to in vitro maturation and subsequent fertilization and in vitro culture in both systems. The structures were evaluated post-maturation and post-culture, for evaluation of nuclear maturation and amount of cells / blastocyst, respectively. No statistical differences (p>0.05) were observed in the different parameters studied. We conclude that both tested systems proved to be efficient for bovine PIVE.
Subject(s)
Animals , Cattle , Embryonic Development , Embryo, Mammalian/embryology , Incubators/veterinary , In Vitro Techniques/methods , In Vitro Techniques/veterinary , Embryo Culture Techniques/methods , Embryo Culture Techniques/veterinary , Reproductive Techniques, Assisted/veterinaryABSTRACT
This study aimed to evaluate three different incubation systems in nuclear maturation of bovine oocytes.Follicles of 2 to 8 mm in diameter were aspirated for obtaining cumulus-oocyte complexes (COCs). Thereafter,COCs were subjected to in vitro maturation (IVM) in TCM-199 medium supplemented at 38.5 °C for 23 h indifferent incubation systems: bench incubator with high oxygen (20% O2 in air), bench incubator with lowoxygen (5% O2) and portable incubator with low oxygen. After IVM, cumulus cells were removed and oocyteswere stained with Hoechst 33342. The slides were analyzed by a fluorescence microscope and the oocytes wereclassified into: degenerated (DG), metaphase I (MI) and metaphase II (MII). No difference (P > 0.05) wasobserved among the rates of DG, MI and MII for the different incubation systems. Therefore, the use of portableincubation system is a viable alternative for in vitro maturation of bovine oocytes.(AU)
Subject(s)
Animals , Female , Cattle , Cattle/genetics , Cattle/physiology , In Vitro Oocyte Maturation Techniques/methods , Infectious Disease Incubation PeriodABSTRACT
tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16 106 and 20 106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16 106 and 20 106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20 106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.(AU)
Subject(s)
Animals , Male , Cattle , Spermatozoa/classification , Spermatozoa/cytology , Cattle/embryology , Embryo, Mammalian , Fertilization in Vitro/veterinaryABSTRACT
The present study investigated the effects of different concentrations of linear crotamine (CrL), used asagent for cell transfection, on in vitro development of bovine embryos. Embryos were exposed to 0; 5 and 12.5µM of CrL in Synthetic Oviductal Fluid (SOF) for 6 h. Vehicle solution was NaCl 150 mM and not exposedembryos (in vitro fertilized-IVF group) was included. In vitro developmental competence was evaluated bycleavage (day 2), blastocyst (day 7 and 8), and hatching (day 8) rates. No difference (P > 0.05) was observedamong CrL groups and control group (IVF) for all observed parameters. In conclusion, the use of the tested CrLconcentrations for 6 h during in vitro culture of bovine embryos have no deleterious effects on embryodevelopment.(AU)
Subject(s)
Animals , Female , Cattle , Cattle/embryology , Embryo, Mammalian , Teratogens , Cell-Penetrating PeptidesABSTRACT
This study aimed to evaluate three different incubation systems in nuclear maturation of bovine oocytes.Follicles of 2 to 8 mm in diameter were aspirated for obtaining cumulus-oocyte complexes (COCs). Thereafter,COCs were subjected to in vitro maturation (IVM) in TCM-199 medium supplemented at 38.5 °C for 23 h indifferent incubation systems: bench incubator with high oxygen (20% O2 in air), bench incubator with lowoxygen (5% O2) and portable incubator with low oxygen. After IVM, cumulus cells were removed and oocyteswere stained with Hoechst 33342. The slides were analyzed by a fluorescence microscope and the oocytes wereclassified into: degenerated (DG), metaphase I (MI) and metaphase II (MII). No difference (P > 0.05) wasobserved among the rates of DG, MI and MII for the different incubation systems. Therefore, the use of portableincubation system is a viable alternative for in vitro maturation of bovine oocytes.
Subject(s)
Female , Animals , Cattle , Cattle/physiology , Cattle/genetics , Infectious Disease Incubation Period , In Vitro Oocyte Maturation Techniques/methodsABSTRACT
The present study investigated the effects of different concentrations of linear crotamine (CrL), used asagent for cell transfection, on in vitro development of bovine embryos. Embryos were exposed to 0; 5 and 12.5µM of CrL in Synthetic Oviductal Fluid (SOF) for 6 h. Vehicle solution was NaCl 150 mM and not exposedembryos (in vitro fertilized-IVF group) was included. In vitro developmental competence was evaluated bycleavage (day 2), blastocyst (day 7 and 8), and hatching (day 8) rates. No difference (P > 0.05) was observedamong CrL groups and control group (IVF) for all observed parameters. In conclusion, the use of the tested CrLconcentrations for 6 h during in vitro culture of bovine embryos have no deleterious effects on embryodevelopment.
Subject(s)
Female , Animals , Cattle , Cattle/embryology , Embryo, Mammalian , Cell-Penetrating Peptides , TeratogensABSTRACT
tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16 106 and 20 106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16 106 and 20 106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20 106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.