关键词: Co-culture Cumulus cells IVM rescue Immature oocytes In vitro maturation

Mesh : Pregnancy Female Humans Coculture Techniques In Vitro Oocyte Maturation Techniques / methods Cumulus Cells / metabolism Fertilization in Vitro / methods Sperm Injections, Intracytoplasmic Prospective Studies Oocytes / metabolism

来  源:   DOI:10.1016/j.morpho.2022.02.002

Abstract:
OBJECTIVE: In stimulated IVF-ICSI cycles, follicles at different stages of maturation can be aspirated during oocyte pickup. Nowadays, only mature oocytes (metaphase 2 stage) are used and immature oocytes (germinal vesicle and metaphase 1 stages), which are judged unfit for fertilization, are non-used at day 0. In our IVF center, the rate of immature oocytes recovered is around 25%. A significant number of this precious resource is therefore non-used every day in IVF laboratories. The objective of our study was to evaluate the competence of our in vitro maturation autologous coculture method on the maturation and developmental potential of immature oocytes obtained from stimulated IVF-ICSI cycles, in order to obtain additional embryos for the couple as a rescue system to increase the changes of cumulative pregnancy.
METHODS: This is a prospective study, carried out in the Reproductive Medicine and Biology Unit of the Amiens-Picardy University Hospital (France). It was included 14 couples, managed in IVF-ICSI in our center, from January to March 2020. Thirty-eight oocytes, identified as immature after cumulus-oocyte complexes (COC) stripping for ICSI, were placed in our in vitro maturation medium with the addition of autologous cumulus cells. Oocytes that had reached the metaphase II stage after a maximum of 36 hours of maturation were microinjected. The fertilization and embryonic development potential of the in vitro matured oocytes were compared to those of 148 in vivo matured \"siblings\" oocytes from the same oocyte retrieval, and then also compared to those of 127 in vivo matured oocytes from different patients (control group).
METHODS: Maturation rate, fertilization rate, early cleavage rate and developmental activity to blastulation rate.
UNASSIGNED: Embryo quality at cleavage and blastocyst stages, blastulation rate, and useful blastulation rate.
RESULTS: No significant difference was found in the main and secondary criteria of the study compared to the \"siblings\" in vivo matured oocytes from the same oocyte retrieval. However, a significant difference was obtained on the rate of early cleavage and useful blastulation when our cohort was compared to mature in vivo oocytes from different patients (control group).
CONCLUSIONS: This study has shown that after incubation in our in vitro maturation autologous cumulus cell co-culture with cumulus-oocyte cells, immature oocytes recovered during stimulated cycles can give rise to competent oocytes, i.e., capable of being fertilized, of cleaving, and of developing into embryos up to the blastocyst stage. Our study therefore seems to be in the direction of a favorable use of these immature oocytes obtained after stimulated IVF-ICSI cycles. The continuation of this study by including a larger number of oocytes is necessary in order to evaluate the real contribution of this technique in routine.
摘要:
目的:在刺激的IVF-ICSI周期中,在卵母细胞拾取过程中,可以抽吸成熟不同阶段的卵泡。如今,仅使用成熟卵母细胞(中期2期)和未成熟卵母细胞(生发囊泡和中期1期),被认为不适合受精,在第0天未使用。在我们的试管婴儿中心,未成熟卵母细胞的回收率约为25%。因此,IVF实验室每天都没有使用大量这种宝贵的资源。我们研究的目的是评估我们的体外成熟自体共培养方法对刺激IVF-ICSI周期获得的未成熟卵母细胞的成熟和发育潜力的能力,以获得额外的胚胎作为救助系统,以增加累积妊娠的变化。
方法:这是一项前瞻性研究,在亚眠-皮卡迪大学医院(法国)的生殖医学和生物学部门进行。其中包括14对夫妇,在我们中心的IVF-ICSI中管理,2020年1月至3月。38个卵母细胞,ICSI卵丘-卵母细胞复合物(COC)剥离后鉴定为未成熟,置于我们的体外成熟培养基中,加入自体卵丘细胞。在最多36小时的成熟后达到中期II阶段的卵母细胞被显微注射。将体外成熟卵母细胞的受精和胚胎发育潜力与来自同一卵母细胞取回的148个体内成熟“兄弟姐妹”卵母细胞的受精和胚胎发育潜力进行了比较,然后与来自不同患者(对照组)的127个体内成熟卵母细胞进行比较。
方法:成熟率,受精率,早期卵裂率和发育活性对囊胚率的影响。
未经证实:卵裂和囊胚阶段的胚胎质量,囊胚率,和有用的囊胚率。
结果:与来自相同卵母细胞的体内成熟卵母细胞相比,在研究的主要和次要标准中没有发现显着差异。然而,当我们的队列与来自不同患者(对照组)的体内成熟卵母细胞进行比较时,在早期卵裂率和有用囊胚形成率上获得了显著差异.
结论:这项研究表明,在我们体外成熟的自体卵丘细胞与卵丘-卵母细胞共培养后,在刺激周期中回收的未成熟卵母细胞可以产生有能力的卵母细胞,即,能够受精,裂开,并发育成胚胎直至胚泡阶段。因此,我们的研究似乎是在刺激IVF-ICSI周期后获得的这些未成熟卵母细胞的有利使用方向。为了评估该技术在常规中的真正贡献,有必要通过包括大量卵母细胞来继续进行这项研究。
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