Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos

ABSTRACT: Spatial and temporal synchrony and compatibility between the receptor oocyte and the donor cell nucleus are necessary for the process of embryo cloning to allow nuclear reprogramming and early embryonic development. The objective of the present study was to evaluate three cell cycle synchr...

Full description

Autores:
Gómez Morales, Natalia Andrea
Ramírez Hernández, Mónica Marcela
Ruiz Cortés, Zulma Tatiana
Tipo de recurso:
Article of investigation
Fecha de publicación:
2018
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
eng
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/32697
Acceso en línea:
https://hdl.handle.net/10495/32697
Palabra clave:
Reprogramación Celular
Cellular Reprogramming
Partenogénesis
Parthenogenesis
Clonación de Organismos
Cloning, Organism
Rights
openAccess
License
http://creativecommons.org/licenses/by/2.5/co/
id UDEA2_ec91d0d8bd178cf7838921547a7205bb
oai_identifier_str oai:bibliotecadigital.udea.edu.co:10495/32697
network_acronym_str UDEA2
network_name_str Repositorio UdeA
repository_id_str
dc.title.spa.fl_str_mv Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
dc.title.alternative.spa.fl_str_mv Sincronização do ciclo celular de fibroblastos primários e efeitos na produção de embriões bovinos clonados por handmade (HMC)
title Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
spellingShingle Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
Reprogramación Celular
Cellular Reprogramming
Partenogénesis
Parthenogenesis
Clonación de Organismos
Cloning, Organism
title_short Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
title_full Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
title_fullStr Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
title_full_unstemmed Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
title_sort Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryos
dc.creator.fl_str_mv Gómez Morales, Natalia Andrea
Ramírez Hernández, Mónica Marcela
Ruiz Cortés, Zulma Tatiana
dc.contributor.author.none.fl_str_mv Gómez Morales, Natalia Andrea
Ramírez Hernández, Mónica Marcela
Ruiz Cortés, Zulma Tatiana
dc.subject.decs.none.fl_str_mv Reprogramación Celular
Cellular Reprogramming
Partenogénesis
Parthenogenesis
Clonación de Organismos
Cloning, Organism
topic Reprogramación Celular
Cellular Reprogramming
Partenogénesis
Parthenogenesis
Clonación de Organismos
Cloning, Organism
description ABSTRACT: Spatial and temporal synchrony and compatibility between the receptor oocyte and the donor cell nucleus are necessary for the process of embryo cloning to allow nuclear reprogramming and early embryonic development. The objective of the present study was to evaluate three cell cycle synchronization methods on a primary bovine fibroblast culture for 24, 48, or 72 h. These fibroblasts were used as nuclear donors to evaluate their in vitro developmental potential and the quality of the embryos produced through handmade cloning (HMC). No differences were found between the methods used for fibroblast synchronization in G0/G1 (p > 0.05). Production of clones from fibroblasts in four groups– no treatment at 0 h and using serum restriction SR, high culture confluence HCC, and SR+HCC at 24 h– resulted in high cleavage rates that were not different. Embryo production rates were 37.9%, 29.5%, and 30.9% in the 0h, SR24h, and SR+HHC24h groups, respectively, and 19.3% in the HCC group, which was significantly different from the other three (p < 0.05). There were no differences in the quality parameter among the clones produced with fibroblasts subjected to the different synchronization. Finally, when overall clone production was compared versus parthenotes and IVF embryos, the only difference was between clones and parthenogenetic embryos with zona pellucida (30.2% vs 38.6%). The number of blastomeres from the blastocytes produced through IVF was significantly greater than those from embryos activated parthenogenetically and from clones (117, 80, 75.9, and 67.1, respectively). The evaluation of three synchronization methods at different time points did not demonstrate an increase in the percentage of fibroblasts in the G0/G1 phases of the cell cycle; however, good quality and high cloning rates were obtained, suggesting that it is not always necessary to subject the cells to any synchronization treatments, as they would yield equally good cloning results.
publishDate 2018
dc.date.issued.none.fl_str_mv 2018
dc.date.accessioned.none.fl_str_mv 2022-12-10T14:59:06Z
dc.date.available.none.fl_str_mv 2022-12-10T14:59:06Z
dc.type.spa.fl_str_mv info:eu-repo/semantics/article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.hasversion.spa.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.coar.spa.fl_str_mv http://purl.org/coar/resource_type/c_2df8fbb1
dc.type.redcol.spa.fl_str_mv https://purl.org/redcol/resource_type/ART
dc.type.local.spa.fl_str_mv Artículo de investigación
format http://purl.org/coar/resource_type/c_2df8fbb1
status_str publishedVersion
dc.identifier.issn.none.fl_str_mv 1518-2797
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/10495/32697
dc.identifier.doi.none.fl_str_mv 10.1590/1809-6891v19e-48555
dc.identifier.eissn.none.fl_str_mv 1809-6891
identifier_str_mv 1518-2797
10.1590/1809-6891v19e-48555
1809-6891
url https://hdl.handle.net/10495/32697
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.ispartofjournalabbrev.spa.fl_str_mv Ciênc. Anim. Bras.
dc.rights.spa.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.uri.*.fl_str_mv http://creativecommons.org/licenses/by/2.5/co/
dc.rights.accessrights.spa.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.creativecommons.spa.fl_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/2.5/co/
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by/4.0/
dc.format.extent.spa.fl_str_mv 17
dc.format.mimetype.spa.fl_str_mv application/pdf
dc.publisher.spa.fl_str_mv Universidade Federal de Goiás, Escola de Veterinária
dc.publisher.group.spa.fl_str_mv Biogénesis
dc.publisher.place.spa.fl_str_mv Goiânia, Brasil
institution Universidad de Antioquia
bitstream.url.fl_str_mv https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/1/GomezNatalia_2018_PrimaryFibroblast.pdf
https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/2/license_rdf
https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/3/license.txt
bitstream.checksum.fl_str_mv 0a983a4720c07c595b4b510c36808e2b
1646d1f6b96dbbbc38035efc9239ac9c
8a4605be74aa9ea9d79846c1fba20a33
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
repository.name.fl_str_mv Repositorio Institucional Universidad de Antioquia
repository.mail.fl_str_mv andres.perez@udea.edu.co
_version_ 1812173151448072192
spelling Gómez Morales, Natalia AndreaRamírez Hernández, Mónica MarcelaRuiz Cortés, Zulma Tatiana2022-12-10T14:59:06Z2022-12-10T14:59:06Z20181518-2797https://hdl.handle.net/10495/3269710.1590/1809-6891v19e-485551809-6891ABSTRACT: Spatial and temporal synchrony and compatibility between the receptor oocyte and the donor cell nucleus are necessary for the process of embryo cloning to allow nuclear reprogramming and early embryonic development. The objective of the present study was to evaluate three cell cycle synchronization methods on a primary bovine fibroblast culture for 24, 48, or 72 h. These fibroblasts were used as nuclear donors to evaluate their in vitro developmental potential and the quality of the embryos produced through handmade cloning (HMC). No differences were found between the methods used for fibroblast synchronization in G0/G1 (p > 0.05). Production of clones from fibroblasts in four groups– no treatment at 0 h and using serum restriction SR, high culture confluence HCC, and SR+HCC at 24 h– resulted in high cleavage rates that were not different. Embryo production rates were 37.9%, 29.5%, and 30.9% in the 0h, SR24h, and SR+HHC24h groups, respectively, and 19.3% in the HCC group, which was significantly different from the other three (p < 0.05). There were no differences in the quality parameter among the clones produced with fibroblasts subjected to the different synchronization. Finally, when overall clone production was compared versus parthenotes and IVF embryos, the only difference was between clones and parthenogenetic embryos with zona pellucida (30.2% vs 38.6%). The number of blastomeres from the blastocytes produced through IVF was significantly greater than those from embryos activated parthenogenetically and from clones (117, 80, 75.9, and 67.1, respectively). The evaluation of three synchronization methods at different time points did not demonstrate an increase in the percentage of fibroblasts in the G0/G1 phases of the cell cycle; however, good quality and high cloning rates were obtained, suggesting that it is not always necessary to subject the cells to any synchronization treatments, as they would yield equally good cloning results.RESUMO: A sincronia espacial e temporal e a compatibilidade entre o oócito receptor e o núcleo celular doador são necessárias para o processo de clonagem de embriões a fim de permitir a reprogramação nuclear e o desenvolvimento embrionário precoce. O objetivo do presente estudo foi avaliar três métodos de sincronização do ciclo celular em uma cultura primária de fibroblastos bovinos durante 24, 48 ou 72 h. Estes fibroblastos foram utilizados como doadores nucleares para avaliar o seu potencial de desenvolvimento in vitro e a qualidade dos embriões produzidos por meio da técnica de Handmade cloning (HMC). Não foram encontradas diferenças entre os métodos utilizados para a sincronização de fibroblastos em G0 / G1 (p> 0,05). Produção de clones de fibroblastos nos quatro grupos – sem tratamento a 0 h e com restrição de soro RS, alta confluência celular ACC e RS + ACC às 24 h – resultou em altas taxas de clivagem que não foram diferentes. As taxas de produção de embriões foram de 37,9%, 29,5% e 30,9% nos grupos 0h, RS24h e RS + ACC24h, respectivamente, e 19,3% no grupo ACC, que foi significativamente diferente dos outros três (p <0,05). Não houve diferenças no parâmetro de qualidade entre os clones produzidos com fibroblastos submetidos à sincronização diferente. Finalmente, quando a produção geral de clones foi comparada versus partenotos e embriões de FIV, a única diferença foi entre clones e embriões partenogênicos com zona pelúcida (30,2% vs 38,6%). O número de blastômeros dos blastocitos produzidos através da FIV foi significativamente maior do que os de embriões ativados partenogeneticamente e de clones (117, 80, 75,9 e 67,1, respectivamente). A avaliação de três métodos de sincronização em diferentes pontos de tempo não demonstrou um aumento na porcentagem de fibroblastos nas fases G0/G1 do ciclo celular. No entanto, obteve-se boa qualidade e altas taxas de clonagem, sugerindo que nem sempre é necessário submeter as células a quaisquer tratamentos de sincronização, uma vez que renderiam resultados de clonagem igualmente bons.COL006656117application/pdfengUniversidade Federal de Goiás, Escola de VeterináriaBiogénesisGoiânia, Brasilinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by/4.0/Primary fibroblast cell cycle synchronization and effects on handmade cloned (HMC) bovine embryosSincronização do ciclo celular de fibroblastos primários e efeitos na produção de embriões bovinos clonados por handmade (HMC)Reprogramación CelularCellular ReprogrammingPartenogénesisParthenogenesisClonación de OrganismosCloning, OrganismCiênc. Anim. Bras.Ciencia Animal Brasileira11719ORIGINALGomezNatalia_2018_PrimaryFibroblast.pdfGomezNatalia_2018_PrimaryFibroblast.pdfArtículo de investigaciónapplication/pdf1298817https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/1/GomezNatalia_2018_PrimaryFibroblast.pdf0a983a4720c07c595b4b510c36808e2bMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8927https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/2/license_rdf1646d1f6b96dbbbc38035efc9239ac9cMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://bibliotecadigital.udea.edu.co/bitstream/10495/32697/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/32697oai:bibliotecadigital.udea.edu.co:10495/326972022-12-10 09:59:06.681Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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