Meiosis Resumption of Immature Human Oocytes following Treatment with Calcium Ionophore In Vitro

(Pages: 109-118)
Elham Fazeli, Ph.D., 1,2Ahmad Hosseini, Ph.D., 2Mohammad-Hasan Heidari, Ph.D., 1,3Fattaneh Farifteh-Nobijari, Ph.D., 3,4Mohammad Salehi, Ph.D., 3,5Hojjat-Allah Abbaszadeh, Ph.D., 1Hamid Nazarian, Ph.D., 1Zahra Shams Mofarahe, Ph.D., 1Saman Ayoubi, M.Sc., 6Sara Hosseini, Ph.D., 3Mona Shayeghpour, M.Sc., 4Mojgan Bandehpour, Ph.D., 3,5,*Marefat Ghaffari Novin, Ph.D., 1,3,*
Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Mehr Fertility Research Center, Guilan University of Medical Sciences, Rasht, Iran
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Genetics and In Vitro Assisted Reproductive (GIVAR) Center, Erfan Hospital, Tehran, Iran
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran Iran
Infertility and Reproductive Health Research Centre, Sara Hospital, Tehran, Iran
Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Mehr Fertility Research Center, Guilan University of Medical Sciences, Rasht, Iran
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Genetics and In Vitro Assisted Reproductive (GIVAR) Center, Erfan Hospital, Tehran, Iran
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran Iran
Infertility and Reproductive Health Research Centre, Sara Hospital, Tehran, Iran
* Corresponding Address: P.O.Box: 1985717443 Cellular and Molecular Biology Research Centre Shahid Beheshti University of Medical Sciences TehranIran P.O.Box: 1985717443 Department of Biology and Anatomical Sciences School of Medicine Shahid Beheshti University of Medical Sciences TehranIran Email: bandehpour@gmail.com, mghaffarin@yahoo.com
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Fazeli Elham, Hosseini Ahmad, Heidari Mohammad-Hasan, Farifteh-Nobijari Fattaneh, Salehi Mohammad, Abbaszadeh Hojjat-Allah, Nazarian Hamid, Shams Mofarahe Zahra, Ayoubi Saman, Hosseini Sara, Shayeghpour Mona, Bandehpour Mojgan, Ghaffari Novin Marefat. Meiosis Resumption of Immature Human Oocytes following Treatment with Calcium Ionophore In Vitro. Cell J. 2021; 23(1): 109-118.

Abstract

Objective:

In vitro maturation (IVM) of human oocytes is used to induce meiosis progression in immature retrieved oocytes. Calcium (Ca2+) has a central role in oocyte physiology. Passage through meiosis phase to another phase is controlled by increasing intracellular Ca2+. Therefore, the current research was conducted to evaluate the role of calcium ionophore (CI) on human oocyte IVM.

Materials and Methods:

In this clinical trial study, immature human oocytes were obtained from 216 intracytoplasmic sperm injection (ICSI) cycles. After ovarian stimulation, germinal vesicle (GV) stage oocytes were collected and categorized into two groups: with and without 10 µM CI treatment. Next, oocyte nuclear maturation was assessed after 24–28 hours of culture. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used to assess the transcript profile of several oocyte maturation-related genes (MAPK3, CCNB1, CDK1, and cyclin D1 [CCND1]) and apoptotic-related genes (BCL-2, BAX, and Caspase-3). Oocyte glutathione (GSH) and reactive oxygen species (ROS) levels were assessed using Cell Tracker Blue and 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent dye staining. Oocyte spindle configuration and chromosome alignment were analysed by immunocytochemistry.

Results:

The metaphase II (MII) oocyte rate was higher in CI‐treated oocytes (73.53%) compared to the control (67.43%) group, but this difference was not statistically significant (P=0.13). The mRNA expression profile of oocyte maturation-related genes (MAPK3, CCNB1, CDK1, and CCND1) (P<0.05) and the anti-apoptotic BCL-2 gene was remarkably up-regulated after treatment with CI (P=0.001). The pro-apoptotic BAX and Caspase-3 relative expression levels did not change significantly. The CI‐treated oocyte cytoplasm had significantly higher GSH and lower ROS (P<0.05). There was no statistically significant difference in meiotic spindle assembly and chromosome alignment between CI treatment and the control group oocytes.

Conclusion:

The finding of the current study supports the role of CI in meiosis resumption of human oocytes. (Registration Number: IRCT20140707018381N4)