Past Issue

Volume 18 , Number 2,Jul-Sep (Summer) 2016, Serial Number: 70, Pages: 205-213

Evaluation of Mouse Oocyte In Vitro Maturation Developmental Competency in Dynamic Culture Systems by Design and Construction of A Lab on A Chip Device and Its Comparison with Conventional Culture System


Behnaz Sadeghzadeh Oskouei, M.Sc, 1, Maryam Pashaiasl, M.D., Ph.D, 1, 2, Mohammad Hasan Heidari, Ph.D, 3, Mohammad Salehi, Ph.D, 4, Hadi Veladi, Ph.D, 5, Firuz Ghaderi Pakdel, Ph.D, 6, Parviz Shahabi, Ph.D, 7, Marefat Ghaffari Novin, M.D., Ph.D, 4, *,
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Womens Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Department of Anatomy and Biology, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Cellular and Molecular Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Department of Electronic Engineering, Tabriz University, Tabriz, Iran
Department of Physiology, Urmia University of Medical Sciences, Urmia, Iran
Department of Physiology, Tabriz University of Medical Sciences, Tabriz, Iran
*Corresponding Address: P.O.Box: 1985717443 Cellular and Molecular Biology Research Centre Shahid Beheshti University of Medical Sciences Tehran Iran Email:mghaffarin@yahoo.com

Abstract

Objective

In conventional assisted reproductive technology (ART), oocytes are cultured in static microdrops within Petri dishes that contain vast amounts of media. However, the in vivo environment is dynamic. This study assesses in vitro oocyte maturation through the use of a new microfluidic device. We evaluate oocyte fertilization to the blastocyct stage and their glutathione (GSH) contents in each experimental group.

Materials and Methods

In this experimental study, we established a dynamic culture condition. Immature oocytes were harvested from ovaries of Naval Medical Research Institute (NMRI) mice. Oocytes were randomly placed in static (passive) and dynamic (active) in vitro maturation (IVM) culture medium for 24 hours. In vitro matured oocytes underwent fertilization, after which we placed the pronucleus (PN) stage embryos in microdrops and followed their developmental stages to blastocyst formation after 3 days. GSH content of the in vitro matured oocytes was assessed by monochlorobimane (MCB) staining.

Results

We observed significantly higher percentages of mature metaphase II oocytes (MII) in the passive and active dynamic culture systems (DCS) compared to the static group (P<0.01). There were significantly less mean numbers of germinal vesicle (GV) and degenerated oocytes in the passive and active dynamic groups compared to the static group (P<0.01). Fertilization and blastocyst formation rate in the dynamic systems were statistically significant compared to the static cultures (P<0.01). There was significantly higher GSH content in dynamically matured oocytes compared to statically matured oocytes (P<0.01).

Conclusion

Dynamic culture for in vitro oocyte maturation improves their developmental competency in comparison with static culture conditions.