Effect of cilostazol on in vitro production of sheep

Detalhes bibliográficos
Autor(a) principal: Silva, Jossimara de Melo
Data de Publicação: 2023
Outros Autores: Freitas, Beatriz Cavalcanti de, Souza, William Matheus, Santos, Damaris Raquel Pires dos, Aureliano, Érika Karoline de Oliveira, Lima, Andreza Mayara Carneiro, Oliveira, Luana Kealy Pimentel de, Cordeiro, Mabel Freitas, Lopes Júnior, Edilson Soares
Tipo de documento: preprint
Idioma: por
Título da fonte: SciELO Preprints
Texto Completo: https://preprints.scielo.org/index.php/scielo/preprint/view/6009
Resumo: The objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacuum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P > 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P > 0.05).
id SCI-1_524e02b3bd3ee80d46d19c708ff4b1d3
oai_identifier_str oai:ops.preprints.scielo.org:preprint/6009
network_acronym_str SCI-1
network_name_str SciELO Preprints
repository_id_str
spelling Effect of cilostazol on in vitro production of sheepEfecto del cilostazol en la producción in vitro de ovejas Efeito do cilostazol na produção in vitro de ovinosembriãoFIVMIVovelhaPIVembryoIVFIVMovineIVPembriónFIVMIVovinoPIVThe objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacuum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P > 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P > 0.05).El objetivo de este estudio fue evaluar el efecto de la inclusión de cilostazol en el medio de maduración in vitro de ovocitos sobre la producción in vitro de embriones ovinos. Para ello, se realizaron recolecciones de ovocitos provenientes de ovarios obtenidos en un matadero mediante el método de aspiración folicular con bomba de vacío. Los ovocitos se dividieron en cuatro grupos de maduración: grupo CON, donde los complejos cúmulus ovocitos se sumergieron en TCM-199, suplementado con 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sodio, 10% (v/v) de suero fetal bovino (SFB), 10 ng/mL de factor de crecimiento epidérmico (EGF), 10 µg/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol y 100 µM de cisteamina; y en los grupos CILO0,3; CILO1 y CILO10, los ovocitos se maduraron en el medio del grupo CON, pero sin la adición de cisteamina y suplementado con las concentraciones de 0,3; 1 y 10 µM, respectivamente. Después de 24 horas, los ovocitos se evaluaron en cuanto a la presencia o no de células del cúmulus y en cuanto al grado de expansión y se destinaron a la fecundación in vitro, en medio FIV, junto con espermatozoides. Después de la FIV, los presuntos cigotos siguieron para el cultivo in vitro. Se evaluaron las clivajes en el día 2, siendo el día 0 el día del inicio del CIV. Los resultados se expresaron en porcentaje y las variables de expansión de las células del cúmulus y número de estructuras clivadas se compararon mediante la prueba del chi-cuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, EE. UU., 2021). Los resultados se consideraron significativos cuando P < 0,05. En relación a la expansión de las células del cúmulus, todos los grupos presentaron el 100% de expansión. No hubo diferencias significativas en cuanto al grado de expansión de las células del cúmulus entre los grupos suplementados con cilostazol y cisteamina (P > 0,05), así como no hubo diferencias significativas entre las tasas de clivaje entre los grupos suplementados con cilostazol y cisteamina (P > 0,05).O objetivo deste estudo foi avaliar o efeito da inclusão de cilostazol no meio de maturação in vitro de oócitos sobre produção in vitro de embriões ovinos. Para isso, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro por meio do método de aspiração folicular com bomba de vácuo. Os oócitos foram divididos em quatro grupos de maturação: grupo CON, onde os complexos cumulus oócitos foram imersos em TCM-199, suplementado com 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sódio, 10% (v/v) de soro fetal bovino (SFB), 10 ng/mL de fator de crescimento epidérmico (EGF), 10 µg/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol e 100 µM de cisteamina; e nos grupos CILO0,3; CILO1 e CILO10, os oócitos foram maturados no meio do grupo CON, mas sem a adição de cisteamina e suplementado com as concentrações de de 0,3; 1 e 10 µM , respectivamente. Após 24h, os oócitos foram avaliados quanto a presença ou não de células do cumulus e quanto ao grau de expansão e destinados à fecundação in vitro, em meio FIV, juntamente com espermatozoides. Após a FIV, os presumíveis zigotos seguiram para o cultivo in vitro. Foram avaliadas clivagens no dia 2, sendo dia 0 o dia do início do CIV. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P < 0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Não houve diferenças significativas quanto ao grau de expansão das células do cumulus entre os grupos suplementados com cilostazol e cisteamina (P > 0,05), assim como não houve diferenças significativas entre as taxas de clivagem entre os grupos suplementados com cilostazol e cisteamina (P > 0,05).SciELO PreprintsSciELO PreprintsSciELO Preprints2023-05-05info:eu-repo/semantics/preprintinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://preprints.scielo.org/index.php/scielo/preprint/view/600910.1590/SciELOPreprints.6009porhttps://preprints.scielo.org/index.php/scielo/article/view/6009/11537Copyright (c) 2023 Jossimara de Melo Silva, Beatriz Cavalcanti de Freitas, William Matheus Souza, Damaris Raquel Pires dos Santos, Érika Karoline de Oliveira Aureliano, Andreza Mayara Carneiro Lima, Luana Kealy Pimentel de Oliveira, Mabel Freitas Cordeiro, Edilson Soares Lopes Júniorhttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessSilva, Jossimara de MeloFreitas, Beatriz Cavalcanti deSouza, William MatheusSantos, Damaris Raquel Pires dosAureliano, Érika Karoline de OliveiraLima, Andreza Mayara CarneiroOliveira, Luana Kealy Pimentel deCordeiro, Mabel FreitasLopes Júnior, Edilson Soaresreponame:SciELO Preprintsinstname:Scientific Electronic Library Online (SCIELO)instacron:SCI2023-05-01T19:53:34Zoai:ops.preprints.scielo.org:preprint/6009Servidor de preprintshttps://preprints.scielo.org/index.php/scieloONGhttps://preprints.scielo.org/index.php/scielo/oaiscielo.submission@scielo.orgopendoar:2023-05-01T19:53:34SciELO Preprints - Scientific Electronic Library Online (SCIELO)false
dc.title.none.fl_str_mv Effect of cilostazol on in vitro production of sheep
Efecto del cilostazol en la producción in vitro de ovejas
Efeito do cilostazol na produção in vitro de ovinos
title Effect of cilostazol on in vitro production of sheep
spellingShingle Effect of cilostazol on in vitro production of sheep
Silva, Jossimara de Melo
embrião
FIV
MIV
ovelha
PIV
embryo
IVF
IVM
ovine
IVP
embrión
FIV
MIV
ovino
PIV
title_short Effect of cilostazol on in vitro production of sheep
title_full Effect of cilostazol on in vitro production of sheep
title_fullStr Effect of cilostazol on in vitro production of sheep
title_full_unstemmed Effect of cilostazol on in vitro production of sheep
title_sort Effect of cilostazol on in vitro production of sheep
author Silva, Jossimara de Melo
author_facet Silva, Jossimara de Melo
Freitas, Beatriz Cavalcanti de
Souza, William Matheus
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
author_role author
author2 Freitas, Beatriz Cavalcanti de
Souza, William Matheus
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Silva, Jossimara de Melo
Freitas, Beatriz Cavalcanti de
Souza, William Matheus
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
dc.subject.por.fl_str_mv embrião
FIV
MIV
ovelha
PIV
embryo
IVF
IVM
ovine
IVP
embrión
FIV
MIV
ovino
PIV
topic embrião
FIV
MIV
ovelha
PIV
embryo
IVF
IVM
ovine
IVP
embrión
FIV
MIV
ovino
PIV
description The objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacuum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P > 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P > 0.05).
publishDate 2023
dc.date.none.fl_str_mv 2023-05-05
dc.type.driver.fl_str_mv info:eu-repo/semantics/preprint
info:eu-repo/semantics/publishedVersion
format preprint
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://preprints.scielo.org/index.php/scielo/preprint/view/6009
10.1590/SciELOPreprints.6009
url https://preprints.scielo.org/index.php/scielo/preprint/view/6009
identifier_str_mv 10.1590/SciELOPreprints.6009
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv https://preprints.scielo.org/index.php/scielo/article/view/6009/11537
dc.rights.driver.fl_str_mv https://creativecommons.org/licenses/by/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv SciELO Preprints
SciELO Preprints
SciELO Preprints
publisher.none.fl_str_mv SciELO Preprints
SciELO Preprints
SciELO Preprints
dc.source.none.fl_str_mv reponame:SciELO Preprints
instname:Scientific Electronic Library Online (SCIELO)
instacron:SCI
instname_str Scientific Electronic Library Online (SCIELO)
instacron_str SCI
institution SCI
reponame_str SciELO Preprints
collection SciELO Preprints
repository.name.fl_str_mv SciELO Preprints - Scientific Electronic Library Online (SCIELO)
repository.mail.fl_str_mv scielo.submission@scielo.org
_version_ 1797047811667132416