Abstract

Platelets contain most of the potent mitogenic factors present in serum and follicular fluid and intraovarian injection of autologous platelet rich plasma (PRP) was shown to improve ovarian function and development of preantral follicles. This study evaluated the effect of PRP on caprine oocyte maturation in vitro and subsequent fertilization and embryonic development. Cumulus oocyte complexes were cultured in a maturation medium supplemented with (1) fetal bovine serum (FBS, control), (2) PRP, extracted from healthy female goats, (3) polyvinyl alcohol (PVA), and (4) PVA plus PRP (PVA-PRP). The degree of cumulus expansion was scored, and denuded oocytes were used for assessment of nuclear maturation, mitochondrial activity, lipid content, redox status, yield and quality of in vitro embryo development, and cryosurvival of the resulting blastocysts. PRP supported the same beneficial effects of FBS on cumulus expansion, nuclear maturation, in vitro developmental competence of oocytes, and survival of vitrified-warmed blastocysts. Moreover, PRP protected oocytes from undesirable effects FBS exerted on the mitochondrial activity and intracytoplasmic lipid content of maturing oocyte. Although PVA could support the same beneficial effects of neither FBS nor PRP on oocyte maturation, its combined addition with PRP improved the yield and quality of oocyte maturation at rates closely similar to PRP. PRP efficiently substitutes beneficial effects of serum during in vitro oocyte maturation and helps maintain the mitochondrial activity of maturing oocytes.

摘要

血小板含有血清和卵泡液中存在的大多数有效的有丝分裂因子，卵巢内注射自体富血小板血浆（PRP）被证明可以改善卵巢功能和窦前卵泡的发育。本研究评估了 PRP 对山羊卵母细胞体外成熟以及随后的受精和胚胎发育的影响。卵丘卵母细胞复合物在补充有 (1) 胎牛血清 (FBS, 对照)、(2) PRP、从健康雌性山羊中提取、(3) 聚乙烯醇 (PVA) 和 (4) PVA 加 PRP 的成熟培养基中培养(PVA-PRP)。对卵丘扩张程度进行评分，并使用裸卵母细胞评估体外核成熟、线粒体活性、脂质含量、氧化还原状态、产量和质量胚胎发育和由此产生的囊胚的冷冻存活。PRP 支持 FBS 对卵丘扩张、核成熟、卵母细胞的体外发育能力和玻璃化加热的胚泡存活的相同有益作用。此外，PRP 保护卵母细胞免受 FBS 对成熟卵母细胞的线粒体活性和胞浆内脂质含量的不良影响。尽管 PVA 可以支持 FBS 和 PRP 对卵母细胞成熟的相同有益效果，但它与 PRP 的联合添加以与 PRP 非常相似的速率提高了卵母细胞成熟的产量和质量。PRP在体外卵母细胞成熟过程中有效替代血清的有益作用，并有助于维持成熟卵母细胞的线粒体活性。