Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Autores
Moro, Lucía Natalia; Hiriart, María Inés; Buemo, Carla Paola; Jarazo, J.; Sestelo, A.; Veraguas, D.; Rodríguez Alvarez, L.; Salamone, Daniel Felipe
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P<0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.
Fil: Moro, Lucía Natalia. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Hiriart, María Inés. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Buemo, Carla Paola. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Sestelo, A.. Jardín Zoológico de Buenos Aires; Argentina
Fil: Veraguas, D.. Universidad de Concepción; Chile
Fil: Rodríguez Alvarez, L.. Universidad de Concepción; Chile
Fil: Salamone, Daniel Felipe. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Materia
Scnt
Aggregation Embryo
Cheetah
Domestic Cat
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/16811
Acceder
id CONICETDig_26141f87fa04ad94760bf0eaaa202851
oai_identifier_str oai:ri.conicet.gov.ar:11336/16811
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expressionMoro, Lucía NataliaHiriart, María InésBuemo, Carla PaolaJarazo, J.Sestelo, A.Veraguas, D.Rodríguez Alvarez, L.Salamone, Daniel FelipeScntAggregation EmbryoCheetahDomestic Cathttps://purl.org/becyt/ford/4.4https://purl.org/becyt/ford/4The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P<0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.Fil: Moro, Lucía Natalia. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Hiriart, María Inés. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Buemo, Carla Paola. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Sestelo, A.. Jardín Zoológico de Buenos Aires; ArgentinaFil: Veraguas, D.. Universidad de Concepción; ChileFil: Rodríguez Alvarez, L.. Universidad de Concepción; ChileFil: Salamone, Daniel Felipe. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaBioscientifica2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/16811Moro, Lucía Natalia; Hiriart, María Inés; Buemo, Carla Paola; Jarazo, J.; Sestelo, A.; et al.; Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression; Bioscientifica; Reproduction; 150; 1; 7-2015; 1-101470-16261741-7899enginfo:eu-repo/semantics/altIdentifier/doi/10.1530/REP-15-0048info:eu-repo/semantics/altIdentifier/url/http://www.reproduction-online.org/content/150/1/1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-11-12T09:56:46Zoai:ri.conicet.gov.ar:11336/16811instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-11-12 09:56:46.573CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
title Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
spellingShingle Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
Moro, Lucía Natalia
Scnt
Aggregation Embryo
Cheetah
Domestic Cat
title_short Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
title_full Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
title_fullStr Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
title_full_unstemmed Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
title_sort Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression
dc.creator.none.fl_str_mv Moro, Lucía Natalia
Hiriart, María Inés
Buemo, Carla Paola
Jarazo, J.
Sestelo, A.
Veraguas, D.
Rodríguez Alvarez, L.
Salamone, Daniel Felipe
author Moro, Lucía Natalia
author_facet Moro, Lucía Natalia
Hiriart, María Inés
Buemo, Carla Paola
Jarazo, J.
Sestelo, A.
Veraguas, D.
Rodríguez Alvarez, L.
Salamone, Daniel Felipe
author_role author
author2 Hiriart, María Inés
Buemo, Carla Paola
Jarazo, J.
Sestelo, A.
Veraguas, D.
Rodríguez Alvarez, L.
Salamone, Daniel Felipe
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Scnt
Aggregation Embryo
Cheetah
Domestic Cat
topic Scnt
Aggregation Embryo
Cheetah
Domestic Cat
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.4
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P<0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.
Fil: Moro, Lucía Natalia. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Hiriart, María Inés. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Buemo, Carla Paola. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Sestelo, A.. Jardín Zoológico de Buenos Aires; Argentina
Fil: Veraguas, D.. Universidad de Concepción; Chile
Fil: Rodríguez Alvarez, L.. Universidad de Concepción; Chile
Fil: Salamone, Daniel Felipe. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
description The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P<0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/16811
Moro, Lucía Natalia; Hiriart, María Inés; Buemo, Carla Paola; Jarazo, J.; Sestelo, A.; et al.; Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression; Bioscientifica; Reproduction; 150; 1; 7-2015; 1-10
1470-1626
1741-7899
url http://hdl.handle.net/11336/16811
identifier_str_mv Moro, Lucía Natalia; Hiriart, María Inés; Buemo, Carla Paola; Jarazo, J.; Sestelo, A.; et al.; Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression; Bioscientifica; Reproduction; 150; 1; 7-2015; 1-10
1470-1626
1741-7899
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1530/REP-15-0048
info:eu-repo/semantics/altIdentifier/url/http://www.reproduction-online.org/content/150/1/1
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Bioscientifica
publisher.none.fl_str_mv Bioscientifica
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.976206

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