Ceratonia siliqua (Carob) extract improved in vitro development of vitrified-warmed mouse germinal vesicle oocytes: asse
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Ceratonia siliqua (Carob) extract improved in vitro development of vitrified-warmed mouse germinal vesicle oocytes: assessment of possible mechanism Azita Faramarzi . Farank Aghaz . Mitra Bakhtiari Zahra Rashidi . Mozafar Khazaei
. Shiva Roshankhah
.
Received: 21 January 2020 / Accepted: 4 October 2020 Ó Springer Nature B.V. 2020
Abstract Oocyte banking is a vital step for safekeeping and spreading genetic resources of animals. It is also used for fertility preservation of human. Oocyte vitrification is closely related to the lower developmental competence which includes the cryo-injury arisen during vitrification. The aim of the present study was to evaluate the maturation, embryonic development and production of reactive oxygen species (ROS) of mice oocytes following the supplementation vitrification media with different concentrations of Ceratonia siliqua (carob) extracts. In this experimental study, germinal vesicle oocytes collected from 8 to 10 week-old female NMRI mice
A. Faramarzi F. Aghaz M. Bakhtiari Z. Rashidi M. Khazaei (&) Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran e-mail: [email protected] A. Faramarzi e-mail: [email protected] F. Aghaz e-mail: [email protected] Z. Rashidi e-mail: [email protected] S. Roshankhah Department of Anatomical Sciences, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran e-mail: [email protected]
(30–40 gr) were randomly divided into six groups of vitrification media supplemented with 0 (control), 5, 10, 20, 30 and 50 lg/ml C. siliqua. After thawing, oocytes were put in an in vitro maturation medium (IVM) (a-MEM: Alpha Minimum Essential Medium). 3–4 and 24 h (hr) later, the oocyte nuclear maturity was checked. Standard in vitro fertilization was performed on the matured oocytes (MII), and embryonic development was followed. Extra- and intracellular ROS was measured in IVM medium after 24 h of oocyte incubation. The addition of 20 and 30 lg/ml C. siliqua extract to vitrification media improved normal morphology of warmed germinal vesicle (GV) oocytes, rate of germinal vesicle break down (GVBD), and metaphase 2 (MII) oocyte formation significantly (p \ 0.05). Fertilization rate, (embryonic development to 2 cells stage, 4–8 cells stage, and [ 8 cells stage increased in the 30 lg/ml C. siliqua group significantly (p \ 0.05). Furthermore, supplementation of 30 lg/ml C. siliqua in vitrification media significantly decreased extra- and intra-cellular of ROS as well as embryonic fragmentation (p \ 0.05). In conclusion, supplementation of GV oocyte vitrification media with carob extract improved maturation, fertilization, and embryonic development rate and decreased extra- and intra-cellular ROS levels. Keywords Oocyte banking Ceratonia siliqua Vitrification Embryo development Reactive oxygen species
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Cell Tissue Bank
Introduction During recent years, oocyte cryopreservation ha
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