P-158 Beneficial effects of platelet rich plasma for human germinal vesicle oocyte in vitro maturation
Authors: Yurchuk T , Piniaiev V , Petrushko M 
Published: June 2022

Abstract

Study question
Can platelet rich plasma (PRP) supplementation to the medium improve outcomes of in vitro maturation of germinal vesicle (GV) oocytes?

Summary answer
PRP as a component of in vitro maturation medium benefits the GV oocyte maturation rate, the mitochondrial membrane potential and the distribution of lipid droplets.

What is known already
The European Society of Human Reproduction and Embryology presents several strategies for maintaining fertility in women, one of which is in vitro maturation (IVM) of oocytes. Despite more than 5,000 children have been born after using the IVM method, the mechanisms involved in meiotic arrest and resumption in oocytes are not fully understood. Hormones and growth factors are components of a complex regulation system which play a mediator role in ovarian function and are involved in meiotic resumption. PRP is a rich source of growth factors and can be effectively used to supplement the medium for in vitro oocyte maturation.

Study design, size, duration
Oocyte maturity was assessed after denudation, and immature gametes at the GV stage (n = 84) were randomly divided into 4 groups for IVM (24 - 48h): 1- without cumulus cells (CC) and PRP, 2 – with PRP without CC, 3 - with CC without PRP, 4 – with CC and PRP. The maturation rate of the oocytes was evaluated in 24 or 48h. The study was carried out between January 2020 to December 2021.

Participants/materials, setting, methods
This prospective study included 30 female patients 33±3.6 years old. Women received exogenous gonadotropins as part of a short protocol of superovulation induction. About 15% oocytes were at the germinal vesicle stage after denudation of retrieved gametes. PRP was prepared by two-step centrifugation method and was added (5%) to the maturation media. Primary endpoints were oocyte maturation rate, mitochondrial membrane potential and lipid droplets distribution evaluated by using JC-1 and Nile Red staining respectively.

Main results and the role of chance
The highest maturation rate was in group 4 (85%) compared with groups 1-3, where the rates were 9.5; 36.4 and 47.6, respectively (p &lt; 0.05). The biggest number of cells with high mitochondrial potential was in oocytes matured with PRP and cumulus cells (4.5 ± 0.7) compared with other groups (1.3 ± 0.3; 3.0 ± 0.5 and 2.7 ± 0.4 for groups 1-3, respectively) (p &lt; 0.05). The distribution of lipid droplets, typical of in vivo matured metaphase II oocytes, was the largest in group 1 (75%) (p &lt; 0.05). There were no differences in this parameter between the other groups.

Limitations, reasons for caution
Limited number of GV oocytes in the study group. Further studies are recommended and should include group of patients with different age range.

Wider implications of the findings
PRP injection into ovarian tissue causes its rejuvenation. Our results confirm for the first time that PRP supplementation to the medium of oocyte in vitro maturation can increase the maturation rate, mitochondrial activity, and improve lipid droplets distribution in human oocytes. These finding are valuable for improvement of IVM medium.

Trial registration number
Not applicable