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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| 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 |
| Aporte de: |
| Sumario: | 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 |
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