Effect of endometrial cell-conditioned medium and platelet-rich plasma on the developmental competence of mouse preantral follicles: An in vitro study / 대한생식의학회지

Objective@#The aim of this study was to evaluate the impacts of platelet-rich plasma (PRP) and conditioned medium (CM) derived from endometrial stromal cells on mouse preantral follicle culture in a two-dimensional system to produce competent mature oocytes for fertilization. @*Methods@#In total, 240 preantral follicles were isolated from female mouse ovarian tissue and divided into four groups. The preantral follicles were isolated three times for each group and then cultured, respectively, in the presence of alpha minimum essential medium (control), PRP, CM, and PRP+CM. The in vitro growth, in vitro maturation, and cleavage percentage of the preantral follicles were investigated. Immunocytochemistry (IHC) was also conducted to monitor the meiotic progression of the oocytes. Additionally, the mRNA expression levels of the two folliculogenesis-related genes (Gdf9 and Bmp15) and two apoptosis-related genes (Bcl2 and Bax) were investigated using real-time polymerase chain reaction. @*Results@#In the PRP, CM, and PRP+CM groups, the preantral follicle maturation (evaluated by identifying polar bodies) were greater than the control group. The cleavage rate in the CM, and PRP+CM groups were also greater than the control group. IHC analysis demonstrated that in each treatment group, meiotic spindle was normal. In the PRP+CM group, the gene expression levels of Bmp15, Gdf9, and Bcl2 were greater than in the other groups. The Bax gene was more strongly expressed in the PRP and control groups than in the other groups. @*Conclusion@#Overall, the present study suggests that the combination of CM and PRP can effectively increase the growth and cleavage rate of mouse preantral follicles in vitro.

comparison of the multiple oocyte maturation gene expression patterns between the newborn and adult mouse ovary

The interaction between follicular cells and oocyte leads to a change in gene expression involved in oocyte maturation processes. The purpose of this study was to quantify the expression of more common genes involved in follicular growth and oocyte developmental competence. In this experimental study, the expression of genes was evaluated with qRT-PCR assay in female BALB/c mice pups at 3-day of pre-pubertal and 8 week old virgin adult ovaries. The tissue was prepared by H and E staining for normal morphological appearance. The data were calculated with the 2-delta Ct formula and assessed using non-parametric two-tailed Mann-Whitney test. The p<0.05 was considered as significant. The data showed a significant increase in the level of Stra8 and GDF9 in adult compared with newborn mice ovaries [p=0.049]. In contrast, a significant decrease in the level of Mvh, REC8, SCP1, SCP3, and ZP2 was observed in adult mice ovaries compared to those in the newborn mice ovaries [all p=0.049 except SCP1: p=0.046]. There was no significant difference in the level of OCT4 and Cx37 expression between adult and newborn mice ovaries. The modifications in gene expression patterns coordinate the follicular developmental processes. Furthermore, the findings showed higher expression level of premeiotic gene [Stra8] and lower level of meiotic entry markers [SCP1, SCP3, and REC8] in juvenile than newborn mouse ovaries