Blastocoel fluid removal and melatonin supplementation in the culture medium improve the viability of vitrified bovine embryos
Autor: | Ligiane O Leme, Elisa Caroline da Silva Santos, Maria Lúcia Gambarini, Carlos Frederico Martins, Benner Geraldo Alves, T. O. Diesel, Thaisa Campos Marques, Heidi Christina Bessler Cumpa, Margot Alves Nunes Dode, Eduardo Barros de Oliveira, Francine Pereira Higino da Costa |
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Rok vydání: | 2020 |
Předmět: |
Fertilization in Vitro
Melatonin Andrology Embryo Culture Techniques 03 medical and health sciences 0302 clinical medicine Human fertilization Food Animals Pregnancy medicine Animals Vitrification Small Animals Cryopreservation 030219 obstetrics & reproductive medicine Zygote Equine Chemistry Hatching Blastocoel 0402 animal and dairy science Embryo 04 agricultural and veterinary sciences 040201 dairy & animal science Blastocyst Apoptosis embryonic structures Dietary Supplements cardiovascular system Animal Science and Zoology Cattle Female medicine.drug |
Zdroj: | Theriogenology. 160 |
ISSN: | 1879-3231 |
Popis: | 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), regardless of vitrification/warming and re-cultivation. The NAC:TNC was smaller for melatonin-treated blastocysts (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. |
Databáze: | OpenAIRE |
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