en
1726-7536
1735-8507
69
2139
288
gregorian
2020
3
17
21
2
online
1
fulltext
en
32500012
The in vitro Analysis of Quality of Ovarian Follicle Culture Systems Using Time-Lapse Microscopy and Quantitative Real-Time PCR
<p>Background: The aim of ovarian follicle <em>in vitro</em> culture is to obtain mature oocytes. To evaluate the efficiency of in vitro culture system, the status of the cultured oocyte can be analyzed. <br />
Methods: The preantral ovarian follicles retrieved from 14-day-old C57Bl/6J mice were cultured in 3D alginate hydrogel. The status of oocytes obtained from mature (3 months old, group A) and immature (3 weeks old, group B) mice was compared to the status of oocytes retrieved from ovarian follicles cultured<em> in vitro</em> (Group C) using qRT-PCR analysis and time-lapse microscopy. In the qRT-PCR analysis, 8 samples for group A (80 oocytes), 8 samples for group B (80 oocytes), and 6 samples for group C (60 oocytes) were included. Time-lapse analysis was performed in group A (oocytes n=31), group B (n=45), and group C (n=21). Statistical analysis was done by Kruskal-Wallis and chi-square tests and differences were considered statistically significant if p<0,05.<br />
Results: The diameter of group C oocytes is lower in comparison to group A oocytes (67 <em>μm vs.</em> 75 <em>μm</em>, correspondingly). Groups B and C oocytes exhibited delayed meiosis in comparison to group A oocytes. Expression levels of six oocyte maturation genes (Ccnb, CDK1, Ccnh, Wee2, Mos and Epab) were evaluated using qRT-PCR analysis. Expression levels of Ccnh and Epab are lowered in group C oocytes compared to the expression levels of these genes in groups A and B oocytes (p<0.05). <br />
Conclusion: Oocytes obtained after ovarian follicles <em>in vitro</em> culture have reduced development competence, future fundamental changes of <em>in vitro</em> culture systems can be expected.</p>
Alginate, <i>In vitro</i> culture, Mice, Oocyte, Ovarian follicle, Real-time PCR, Time-lapse microscopy
094
107
https://www.jri.ir/article/60075
https://www.jri.ir/documents/fullpaper/en/60075.pdf
MaximFilatovFaculty of Biology, Lomonosov Moscow State University, Moscow, Russiamaxfilat@yandex.ru62025
DenisNikishinN.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia62026
YuliaKhramovaFaculty of Biology, Lomonosov Moscow State University, Moscow, Russia62027
MariaSemenovaFaculty of Biology, Lomonosov Moscow State University, Moscow, Russia62028