Blastocoel fluid removal and melatonin supplementation in the culture medium improve the viability of vitrified bovine embryos.

Autor: Marques TC; Federal Institute of Goias, Campus Rio Verde, Rod. Sul Goiana Km 01, Zona Rural, CEP 75901-970, Rio Verde, Goias, Brazil., Santos ECDS; Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Rodovia Goiânia - Nova Veneza, km 8, Campus Samambaia, CEP 74690-900, Goiânia, Goiás, Brazil., Diesel TO; Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Rodovia Goiânia - Nova Veneza, km 8, Campus Samambaia, CEP 74690-900, Goiânia, Goiás, Brazil., Martins CF; Embrapa Cerrados, Br 020, Km 18, CEP 70770-901, Brasília, DF, Brazil., Cumpa HCB; Embrapa Cerrados, Br 020, Km 18, CEP 70770-901, Brasília, DF, Brazil., Leme LO; Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Parque Estação Biológica, PqEB, Av. W5 Norte (final) Caixa Postal 02372, CEP 70770-917, Brasília, DF, Brazil., Dode MAN; Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Parque Estação Biológica, PqEB, Av. W5 Norte (final) Caixa Postal 02372, CEP 70770-917, Brasília, DF, Brazil., Alves BG; Special Academic Unit of Agrarian Sciences, Federal University of Jataí, R. Riachuelo s/n, CEP 75804-020, Jataí, Goias, Brazil., Costa FPH; Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Rodovia Goiânia - Nova Veneza, km 8, Campus Samambaia, CEP 74690-900, Goiânia, Goiás, Brazil., Oliveira EB; Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Rodovia Goiânia - Nova Veneza, km 8, Campus Samambaia, CEP 74690-900, Goiânia, Goiás, Brazil., Gambarini ML; Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Rodovia Goiânia - Nova Veneza, km 8, Campus Samambaia, CEP 74690-900, Goiânia, Goiás, Brazil. Electronic address: mlgambarini@pq.cnpq.br.
Jazyk: angličtina
Zdroj: Theriogenology [Theriogenology] 2021 Jan 15; Vol. 160, pp. 134-141. Date of Electronic Publication: 2020 Nov 04.
DOI: 10.1016/j.theriogenology.2020.10.028
Abstrakt: In this study, we investigated the effects of melatonin supplementation in the culture medium and blastocoel fluid removal (BFR) before vitrification on the quality and viability of in vitro-derived bovine embryos. After fertilization, presumptive zygotes were assigned to one of the following treatments: control, in vitro standard culture (IVC) medium; IVC + M10 -9 , IVC medium supplemented 10 -9  M melatonin; or IVC + M10 -9 BFR, IVC medium supplemented with 10 -9  M melatonin plus BFR on day 7 (D7) of culture. D7 blastocysts were vitrified by the Cryotop method and, after 5 mo of storage, were warmed and incubated for an additional 72 h. The re-expansion rate was evaluated after 2 and 24 h, and the hatching rate was evaluated after 24, 48, and 72 h. At 72 h, the total number of cells (TNC); number of apoptotic cells (NAC); and expression of genes related to oxidative stress (HSPA5), cell metabolism (SLC2A3), cell repair (MSH6), placentation (KRT8 and PLAC8), and implantation (FOSL1) were assessed in the blastocysts. Less than 30% of the control blastocysts re-expanded until 2 h, whereas more than 85% of the IVC + M10 -9 and IVC + M10 -9 BFR blastocysts re-expanded (P < 0.05). The hatching rate of IVC + M10 -9 BFR blastocysts increased at all time points (P < 0.05), reaching 66.8% at 72 h of incubation. The TNC was similar among treatments (P > 0.05), regardless of vitrification/warming and re-cultivation. The NAC:TNC was smaller for melatonin-treated blastocysts (P < 0.05). BFR increased HSPA5 (P = 0.0118) expression and did not affect SLC2A3, MSH6, KRT8, and FOSL1 expression (P > 0.05). In conclusion, melatonin (10 -9  M) supplementation in the culture medium and BFR on D7 of culture increased the hatching rate 24, 48, and 72 h after warming of the vitrified embryos, indicating an improvement in cryotolerance.
(Copyright © 2020. Published by Elsevier Inc.)
Databáze: MEDLINE
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