Document Type : Original Article
Authors
1
Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran
2
Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran.
3
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
4
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
5
Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
6
Genetics and Fertility Unit, Erfan Hospital, Tehran, Iran
7
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
8
Department of Advanced Medical Sciences and Technologies, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Abstract
Background: Testicular cell conditioned medium (TCCM) has been shown to induce female germ cell development
in vitro from embryonic stem cells (ESCs). Testicular cells (TCs) secrete a variety of growth factors such as growth
differentiation factor-9 (GDF-9), bone morphogenetic protein 4 (BMP-4), stem cell factor (SCF), leukemia inhibitory
factor (LIF), and other, that could improve oocyte maturation. Here we have investigated the effect of human TCCM
(hTCCM) on in vitro maturation (IVM) and morphology of mouse oocytes.
Materials and Methods: In this experimental study, 360 germinal vesicle (GV) oocytes were obtained from NMRI
mice, aged 4-6 weeks that had received 5 IU pregnant mare's serum gonadotropin (PMSG) 48 hours before. GV
oocytes were subjected to IVM. 120 GV oocytes were cultured in each medium; hTCCM as the test group, DMEM
+ 20%FBS as the control group and Ham’s F10 + HFF medium as the sham group. The rates of the IVM and perivitelline
space (PVS) changes were recorded at 8, 16 and 24 hours after culture. The metaphase II (MII) oocytes were
subjected for in vitro fertilization (IVF) and the fertilization rate was evaluated after 1, 2, and 3 days.
Results: There was a significant difference between the maturation rates in hTCCM (31.67% MII) and the control [0% MII,
p <0.05, (7.5% MI, 52.5% deg. and 40%GV)] groups but there was not a significant difference between the maturation rates
in hTCCM and the sham group (53.33% MII, P>0.05). IVF success rate for MII oocytes obtained from IVM in the hTCCM
group was 28.94% (n=11). Our data showed that hTCCM is an effective medium for GV oocyte growth and maturation
compared to the control medium.
Conclusion: Our findings show that TCCM supports oocyte IVM in mice and affect oocyte morphology.
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