scholarly journals Un sistema de co-cultivo con células del cumulus oophurus mejora la calidad de embriones bovinos producidos in vitro

2020 ◽  
Vol 31 (3) ◽  
pp. e18181
Author(s):  
Susana Guevara-Chacón ◽  
Hurley Abel Quispe-Ccasa ◽  
José Américo Saucedo-Uriarte ◽  
Ilse Silvia Cayo-Colca
Keyword(s):  
Cumulus Oophorus

El estudio tuvo como objetivo determinar la influencia de un sistema de co-cultivo con células del cumulus oophorus sobre la calidad de embriones bovinos producidos in vitro en función a su viabilidad. Se utilizaron ovarios de bovinos hembra cruzadas, sacrificadas en el Centro de Beneficio Municipal de Chachapoyas, Amazonas, Perú. Los ovocitos aspirados de folículos de 2-6 mm fueron madurados en medio TCM-199 por 24 horas, en una atmósfera humidificada con 6% de CO2 y a 38.5 °C. El estado de maduración fue determinado mediante el estadio nuclear a través de tinción aceto-orceína. Los ovocitos maduros se fertilizaron in vitro con semen congelado de toros Angus durante 18 horas. Los presuntos cigotos fueron distribuidos aleatoriamente para su desarrollo en un grupo en medio con co-cultivo y otro en medio sin co-cultivo (control). Se determinó la calidad embrionaria en función a la actividad enzimática y daño al ADN de los embriones al séptimo día del cultivo, mediante los fluorocromos diacetato de fluoresceína (FDA) y yoduro de propidio (PI), respectivamente. No se encontró asociación significativa (p>0.05) en la cantidad de embriones producidos in vitro, pero hubo diferencia significativa en viabilidad embrionaria a favor del sistema con co-cultivo (p<0.05). El uso de co-cultivos con células de cumulus oophurus mejora la viabilidad y, por lo tanto, la calidad embrionaria, sin influir en la tasa de producción de embriones in vitro.

ÍNDICE DE RECUPERAÇÃO DE OÓCITOS BOVINOS OBTIDOS DE OVÁRIOS ADQUIRIDOS EM ABATEDOURO COMO CONTRIBUIÇÃO PARA OS ESTUDOS DE FECUNDAÇÃO E PRODUÇÃO DE EMBRIÕES IN VITRO

2009 ◽  
Vol 1 (1) ◽  
Author(s):  
Elzylene Léga ◽  
Juliana Maria Avanci Agostinho
Keyword(s):  
Cumulus Oophorus

A produção in vitro de embriões é uma biotécnica utilizada, alternativamente, para acelerar a produção de animais geneticamente superiores além de permitir o aprofundamento dos conhecimentos relativos aos processos fisiológicos, bioquímicos e biotecnológicos da espécie animal de interesse. O desenvolvimento de técnicas in vitro para a produção de embriões de animais domésticos tem grande potencial tanto para pesquisa básica quanto para aplicações a campo e tem sido muito utilizadas e difundidas no Brasil e no mundo. O presente trabalho objetivou classificar oócitos bovinos obtidos de ovários adquiridos em abatedouro, determinando o índice de recuperação dessas estruturas e identificando os de maior viabilidade para o procedimento de fertilização in vitro. Através da aspiração folicular, foram obtidos oócitos posteriormente classificados em escala de 1 a 4, de acordo com a quantidade de camadas de células do complexo cumulus oophorus que envolvem o oócito. Observouse 15,22% de oócitos de qualidade 1; 25,88% de qualidade 2; 35,53% de qualidade 3 e 23,35% de qualidade 4, determinando maiores índices de oócitos recuperados classificados na qualidade 2 e 3, sendo estes fisiologicamente viáveis e capacitados para entrarem em um programa de fertilização in vitro.

Significance of cumulus oophorus in in-vitro fertilization and oocyte viability and fertility

1990 ◽  
Vol 5 (7) ◽  
pp. 847-852 ◽  
Author(s):  
S. Magier ◽  
H.H. van der Ven ◽  
K. Diedrich ◽  
D. Krebs
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Cumulus Oophorus

scholarly journals Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro

Reproduction ◽  
2012 ◽  
Vol 144 (6) ◽  
pp. 687-697 ◽  
Author(s):  
J Beek ◽  
H Nauwynck ◽  
D Maes ◽  
A Van Soom
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Inhibitory Effect ◽  
Fertilization Rate ◽  
Quality Parameters ◽  
Fertilization In Vitro ◽  
Cumulus Oophorus ◽  
Sperm Penetration

In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 μM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1–TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

scholarly journals CAPACITATION OF MOUSE SPERMATOZOA IN VITRO: INVOLVEMENT OF EPIDIDYMAL SECRETIONS AND CUMULUS OOPHORUS

Reproduction ◽  
1974 ◽  
Vol 41 (1) ◽  
pp. 253-256 ◽  
Author(s):  
R. B. L. GWATKIN ◽  
O. F. ANDERSEN ◽  
D. T. WILLIAMS
Keyword(s):  
Cumulus Oophorus

scholarly journals Assessment of in Vitro Developmental Capacity of Porcine Nuclear-Transferred Embryos Reconstituted with Cumulus Oophorus Cells Undergoing Vital Diagnostics for Apoptosis Detection / Ocena zdolności rozwojowych in vitro klonalnych zarodków świni rekonstytuowanych z jąder komórek wzgórka jajonośnego poddawanych przyżyciowej diagnostyce w kierunku wykrywania apoptozy

2013 ◽  
Vol 13 (3) ◽  
pp. 513-529 ◽  
Author(s):  
Marcin Samiec ◽  
Maria Skrzyszowska
Keyword(s):  
Plasma Membranes ◽  
Dermal Fibroblast ◽  
Annexin V ◽  
Cumulus Cells ◽  
Fibroblast Cells ◽  
Developmental Potential ◽  
Cumulus Oophorus ◽  
Group I ◽  
Cloned Pig

Abstract The objective of the current investigation was to extensively compare the in vitro developmental capabilities between cloned pig embryos reconstructed with the cell nuclei of either cumulus oophorus cells or adult dermal fibroblast cells that were both evaluated as non-apoptotic on the basis of YO-PRO-1- and Annexin V-eGFP-mediated vital analysis for programmed cell suicide. In Group I, the competences of nuclear-transferred (NT) embryos that were derived from non-apoptotic/ non-necrotic (i.e., YO-PRO-1- and Annexin V-eGFP-negative) cumulus cells to complete their development to the morula and blastocyst stages were maintained at the proportions of 155/364 (42.6%) and 54/364 (14.8%), respectively. In Group II, NT embryos that were reconstituted with non-apoptotic and/or non-necrotic adult cutaneous fibroblast cells developed to the morula and blastocyst stages at the rates of 207/358 (57.8%) and 110/358 (30.7%), respectively. Although the in vitro developmental potential of porcine NT embryos derived from non-apoptotic/non-necrotic cumulus cells was significantly lower (P<0.001) than that of NT embryos reconstructed with adult dermal fibroblast cells, the obtained morula/blastocyst formation rates turned out to be considerably higher as compared to the rates reported by other investigators. Altogether, to our knowledge, the comprehensive research aimed at the determination of preimplantation developmental outcomes of cloned pig embryos produced using nuclear donor somatic cells of different provenance (cumulus oophorus cells or adult cutaneous fibroblast cells) that were vitally diagnosed for the lack of proapoptotic transformations in their plasma membranes has not yet been accomplished.

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