Universidad de Buenos Aires, Facultad de...
The efficiency of the in vitro production process (IVP) in mammals is affected by various variables associated with culture conditions during maturation, fertilization and embryonic development. Currently, the percentages of pig blastocysts obtained from\nslaughterhouse ovaries are low compared to o...
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| Otros Autores: | |
| Formato: | Tesis doctoral acceptedVersion |
| Lenguaje: | Español |
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Universidad de Buenos Aires. Facultad de Ciencias Veterinarias
2022
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| Acceso en línea: | http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avaposgra&cl=CL1&d=HWA_6859 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avaposgra/index/assoc/HWA_6859.dir/6859.PDF |
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I28-R145-HWA_6859 |
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dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-145 |
| collection |
Repositorio Digital de la Universidad de Buenos Aires (UBA) |
| language |
Español |
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spa |
| topic |
Celulas In vitro Embriones Fecundación Ciencias Veterinarias |
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Celulas In vitro Embriones Fecundación Ciencias Veterinarias Lorenzo, Maria Soledad Universidad de Buenos Aires, Facultad de... |
| topic_facet |
Celulas In vitro Embriones Fecundación Ciencias Veterinarias |
| description |
The efficiency of the in vitro production process (IVP) in mammals is affected by various variables associated with culture conditions during maturation, fertilization and embryonic development. Currently, the percentages of pig blastocysts obtained from\nslaughterhouse ovaries are low compared to other species. The embryos obtained are of lower quality because the in vitro embryo culture conditions are suboptimal in relation\nto the in vivo environment in which these processes occur. The co-cultured of gametes and embryos with somatic cells is a tool that was developed to improve the in vitro culture\nconditions. These cells can remove toxic or harmful components, protect against\noxidative stress, physically and chemically modify the medium, and secrete growth factors.\nThe objective of this work was to evaluate the effect of co-culture porcine\nembryos produced in vitro with porcine oviductal epithelial cells (POEC) on the in vitro embryonic development and the quality of the embryos obtained. Firstly, a pure culture of POEC capable of being used in co-culture systems was obtained. The epithelial character of the cells in primary culture and passage 1 was confirmed by the expression of E cadherin and cytokeratin and the presence of cilia in the spheroidal vesicles (EV) of\nPOEC. According to the characterization, these cells presented lipid droplets,steroidogenic activity, and the secretion of glycoproteins. The expression pattern of\nhormone receptors coincided with diestrous oviducts and differed between the primary\nculture and passage 1. Secondly, different POEC co-culture systems were used with in in vitro-produced pig embryos. A successful POEC co-culture system was established without adding fetal bovine serum as a supplement. The co culture of passage 1-POEC in monolayers with pig embryos obtained in vitro during the first two days of culture favoured the percentage of blastocysts and their hatching. Although no treatment modified the number of cells in the blastocysts and the apoptosis evaluated by TUNEL, the co-culture with passage 1-POEC decreased reactive oxygen species (ROS) levels in the cleaved porcine embryos obtained in vitro. Co-culture with EV of POEC did not\nrepresent a benefit over embryo culture in traditional media. This work constitutes the\nfirst report on evaluating the quality of porcine embryos produced by IVF in co-culture systems and ROS levels in cleaved porcine embryos obtained by IVF.\nThese results contribute to improving the techniques currently used in the in vitro production of porcine embryos and the evaluation of embryonic quality. Likewise, the\ndeveloped co-culture models could be used to understand the biochemical and\nmolecular mechanisms involved in the maternal-embryonic interaction in the oviductal environment.\n |
| author2 |
Lombardo, Daniel |
| author_facet |
Lombardo, Daniel Lorenzo, Maria Soledad |
| format |
Tesis doctoral Tesis doctoral acceptedVersion |
| author |
Lorenzo, Maria Soledad |
| author_sort |
Lorenzo, Maria Soledad |
| title |
Universidad de Buenos Aires, Facultad de... |
| title_short |
Universidad de Buenos Aires, Facultad de... |
| title_full |
Universidad de Buenos Aires, Facultad de... |
| title_fullStr |
Universidad de Buenos Aires, Facultad de... |
| title_full_unstemmed |
Universidad de Buenos Aires, Facultad de... |
| title_sort |
universidad de buenos aires, facultad de... |
| publisher |
Universidad de Buenos Aires. Facultad de Ciencias Veterinarias |
| publishDate |
2022 |
| url |
http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avaposgra&cl=CL1&d=HWA_6859 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avaposgra/index/assoc/HWA_6859.dir/6859.PDF |
| work_keys_str_mv |
AT lorenzomariasoledad universidaddebuenosairesfacultadde AT lorenzomariasoledad efectosdelcocultivodecelulasepitelialesoviductalesporcinassobrelacalidadydesarrollodeembrionesporcinosproducidosinvitro |
| _version_ |
1824356457214115840 |
| spelling |
I28-R145-HWA_68592024-08-27 Universidad de Buenos Aires, Facultad de... The efficiency of the in vitro production process (IVP) in mammals is affected by various variables associated with culture conditions during maturation, fertilization and embryonic development. Currently, the percentages of pig blastocysts obtained from\nslaughterhouse ovaries are low compared to other species. The embryos obtained are of lower quality because the in vitro embryo culture conditions are suboptimal in relation\nto the in vivo environment in which these processes occur. The co-cultured of gametes and embryos with somatic cells is a tool that was developed to improve the in vitro culture\nconditions. These cells can remove toxic or harmful components, protect against\noxidative stress, physically and chemically modify the medium, and secrete growth factors.\nThe objective of this work was to evaluate the effect of co-culture porcine\nembryos produced in vitro with porcine oviductal epithelial cells (POEC) on the in vitro embryonic development and the quality of the embryos obtained. Firstly, a pure culture of POEC capable of being used in co-culture systems was obtained. The epithelial character of the cells in primary culture and passage 1 was confirmed by the expression of E cadherin and cytokeratin and the presence of cilia in the spheroidal vesicles (EV) of\nPOEC. According to the characterization, these cells presented lipid droplets,steroidogenic activity, and the secretion of glycoproteins. The expression pattern of\nhormone receptors coincided with diestrous oviducts and differed between the primary\nculture and passage 1. Secondly, different POEC co-culture systems were used with in in vitro-produced pig embryos. A successful POEC co-culture system was established without adding fetal bovine serum as a supplement. The co culture of passage 1-POEC in monolayers with pig embryos obtained in vitro during the first two days of culture favoured the percentage of blastocysts and their hatching. Although no treatment modified the number of cells in the blastocysts and the apoptosis evaluated by TUNEL, the co-culture with passage 1-POEC decreased reactive oxygen species (ROS) levels in the cleaved porcine embryos obtained in vitro. Co-culture with EV of POEC did not\nrepresent a benefit over embryo culture in traditional media. This work constitutes the\nfirst report on evaluating the quality of porcine embryos produced by IVF in co-culture systems and ROS levels in cleaved porcine embryos obtained by IVF.\nThese results contribute to improving the techniques currently used in the in vitro production of porcine embryos and the evaluation of embryonic quality. Likewise, the\ndeveloped co-culture models could be used to understand the biochemical and\nmolecular mechanisms involved in the maternal-embryonic interaction in the oviductal environment.\n Fil: Lorenzo, Maria Soledad. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Buenos Aires, Argentina Lombardo, Daniel Lorenzo, Maria Soledad 2022-04-04 La eficiencia del proceso de producción in vitro (PIV) en los mamíferos es\nafectada por diversas variables asociadas a las condiciones de cultivo durante la maduración, fecundación y desarrollo embrionario. Actualmente, los porcentajes de\nblastocistos porcinos obtenidos a partir de ovarios de faena son bajos en comparación\ncon otras especies y los embriones obtenidos de menor calidad, debido a que las condiciones de cultivo embrionario in vitro resultan subóptimas en relación con el\nambiente in vivo en el cual se desarrollan estos procesos. El cocultivo de gametas y embriones con células somáticas es una herramienta que se desarrolló para mejorar las\ncondiciones de cultivo in vitro. Estas células pueden remover componentes tóxicos o\ndeletéreos, proteger del estrés oxidativo, modificar físico-químicamente el medio y secretar factores de crecimiento.\nEl objetivo de este trabajo fue evaluar el efecto del cocultivo de embriones\nporcinos producidos in vitro con células epiteliales oviductales porcinas (CEOP) sobre\nel desarrollo embrionario in vitro y la calidad de los embriones obtenidos. Se decidió utilizar un cocultivo corto, durante los primeros días de desarrollo. En una primera instancia, se obtuvo un cultivo en pureza de CEOP capaz de ser utilizado en sistemas\nde cocultivo. El carácter epitelial en las células en cultivo primario y de pasaje 1 se confirmó por la expresión de cadherina E, citoqueratina y la presencia de cilios en las\nvesículas esferoidales (VE) de CEOP. Según la caracterización, estas células presentaron gotas lipídicas, actividad esteroideogenica, así como también la secreción\nde glucoproteínas como la glucoproteína oviductal 1 (OVGP-1). El patrón de expresión de receptores hormonales coincidió con la utilización de oviductos de diestro, pero difirió\nentre el cultivo primario y el pasaje 1. En una segunda instancia, se utilizaron diferentes\nsistemas de cocultivo de CEOP con embriones porcinos producidos in vitro. Se estableció un sistema de cocultivo con CEOP exitoso, sin el agregado de suero fetal\nbovino como complemento. El cocultivo de CEOP de pasaje 1 en monocapa con embriones porcinos obtenidos in vitro durante los dos primeros días de cultivo favoreció\nel porcentaje de blastocistos y su eclosión. Si bien ningún tratamiento modificó el número de células en los blastocistos y la apoptosis evaluada por TUNEL, el cocultivo\ncon CEOP de pasaje 1 disminuyó los niveles de especies reactivas derivadas del oxígeno (ERO) en los embriones porcinos segmentados, obtenidos in vitro. El cocultivo con VE de CEOP no representó un beneficio con respecto al cultivo de embriones en medios tradicionales.\nEste trabajo constituye el primer reporte de la evaluación de la calidad de embriones porcinos producidos por FIV en sistemas de cocultivo y de la evaluación de\nlos niveles de ERO en embriones porcinos segmentados obtenidos por FIV. Los resultados obtenidos contribuyen a mejorar las técnicas actualmente empleadas en la producción in vitro de embriones porcinos y en la evaluación de la calidad embrionaria.\nAsimismo, los modelos de cocultivo desarrollados podrían ser utilizados para\ncomprender los mecanismos bioquímicos y moleculares implicados en la interacción\nmaterno-embrionaria en el ambiente oviductal.\n application/pdf spa Universidad de Buenos Aires. Facultad de Ciencias Veterinarias info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nend/2.5/ar Celulas In vitro Embriones Fecundación Ciencias Veterinarias Doctora de la Universidad de Buenos Aires en Ciencias Veterinarias Efectos del cocultivo de células epiteliales oviductales porcinas sobre la calidad y desarrollo de embriones porcinos producidos in vitro info:eu-repo/semantics/doctoralThesis info:ar-repo/semantics/tesis doctoral info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avaposgra&cl=CL1&d=HWA_6859 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avaposgra/index/assoc/HWA_6859.dir/6859.PDF |