eprintid: 783
rev_number: 10
eprint_status: archive
userid: 5
dir: disk0/00/00/07/83
datestamp: 2023-10-17 15:38:41
lastmod: 2023-10-17 15:38:41
status_changed: 2023-10-17 15:38:41
type: article
metadata_visibility: show
creators_name: Kandil, O.M.
creators_name: Aboelwafa, F.B.
creators_name: Ismail, E.A.
creators_name: Kandeel, S.M.
creators_name: Ghanem, N.
creators_name: El-Din, A.E.-K.G.
title: Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes
ispublished: pub
subjects: SF
divisions: j13
full_text_status: public
keywords: ascorbic acid; bone morphogenetic protein 15; copper zinc superoxide dismutase; dimethyl sulfoxide; ethylene glycol; gentamicin; growth differentiation factor 9; nitrogen, animal cell; Article; BMP15 gene; controlled study; DNA synthesis; dromedary; female; GDF9 gene; gene expression; gene expression profiling; in vitro oocyte maturation; in vitro study; morphology; nonhuman; oocyte; oocyte maturation; oocyte transport; oocyte viability; oocyte vitrification; real time polymerase chain reaction; RNA extraction; SOD1 gene; supplementation; thawing; upregulation
note: Department of Animal Reproduction and Artificial Insemination, Veterinary Research Institute National Research Centre, Dokki, Cairo, Egypt; Director and supervisor of Accredited (ISO 17025) Embryo and Genetic Resources Conservation Bank in National Research Centre (NRC), Dokki, Cairo, Egypt; Genetics Department, Faculty of Agriculture, Cairo University, Giza, Egypt; Department of Chemistry of Natural Compounds, Institute of Pharmaceutical and Drug Industry, National Research Centre, Dokki, Cairo, Egypt; Animal Production Department, Faculty of Agriculture, Cairo University, Giza, Egypt
abstract: In vitro embryo generation, cryopreservation, and embryo transfer are examples of assisted reproductive technologies that can be used to improve camel genetic performance and fertility. The aim of this study was to investigate the impact of ascorbic acid supplementation to in vitro maturation media on the maturation rate, morphology, and gene expression of fresh and vitrified in vitro matured dromedary camel oocytes. In the current study, 810 oocytes of excellent and good quality were in vitro matured in maturation medium (TCM-199 + 10 ug/ml follicle stimulated hormone + 10 fetal calf serum + 100 IU/ml Pregnant mare serum + 50 μg/ml gentamycin) without any additives to act as a control group (C) and with 50 μg/ml ascorbic acid group (AA) and incubation in a CO2 incubator (38.5 �C, 5 CO2, 20 O2 and 95 humidity) for 40 hours. In vitro matured dromedary camel oocytes with the first polar body (n = 210) in C group and AA group (n = 250) were placed in basic medium (BM) and then placed in vitrification solution1 (VS1) for one minute, followed by the transfer of oocytes to VS2 (double concentration of VS1, containing 20 Ethyl Glycol (EG) and +20 Dimethyl sulfoxide) for 30 seconds. Oocytes were then loaded into sterile 0.25 ml straws and stored in liquid nitrogen for 7-10 days. The normal fresh and vitrified /thawed in vitro matured dromedary camel oocytes were kept in RNA later at a -80°C freezer for gene expression analysis. The maturation rate of dromedary camel oocytes in the in vitro matured AA group was significantly higher than that of the C group. The percentage of normally recovered vitrified/thawed oocytes was higher in the in vitro matured with ascorbic acid (VAA) than in the control (VC) group. The expression pattern of the SOD1 gene and GDF9 gene was upregulated in fresh AA and VAA groups than in the fresh C and VC groups. The profile of the SOD1 gene was more abundant in the vitrified/thawed oocytes VAA group than in the VC group. All vitrified/thawed groups, whether control or ascorbic acid supplemented, had lower levels of SOD1, GDF9, and BMP15 expression, compared to the fresh groups. In conclusion, the supplementation of the maturation medium with ascorbic acid has an increased maturation rate, and normal morphology of vitrified/ thawed oocytes which was linked with upregulation of SOD1, GDF9 genes expression. © 2022, World''s Veterinary Journal. All Rights Reserved.
date: 2022-12-25
publication: World's Veterinary Journal
volume: 12
number: 4
publisher: Scienceline Publication, Ltd
pagerange: 418-429
id_number: 10.54203/scil.2022.wvj52
refereed: TRUE
issn: 2322-4568
official_url: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85161535695&doi=10.54203%2fscil.2022.wvj52&partnerID=40&md5=a388240f6bfe0b8ab8714080dbdb1cc3
j_index: scopus
citation:    (2022)     Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes.   World's Veterinary Journal.  pp. 418-429.  ISSN 2322-4568   
document_url: http://eprints.science-line.com/id/eprint/783/1/article/74/WVJ%2012%284%29%20418-429%2C%20December%2025%2C%202022.pdf