{"id":"hum0490","version":1,"url":"https://humandbs.dbcls.jp/research/hum0490/v1","datePublished":"2024-12-18","versions":[{"version":1,"datePublished":"2024-12-18"}],"title":{"ja":"Progestin-primed ovarian stimulation（PPOS）法を用いた調節卵巣刺激下でプロゲスチン製剤が卵胞発育・卵子の成熟過程に及ぼす影響に関する検討","en":"An investigation into the effects of progestin on follicular development and oocyte maturation during controlled ovarian stimulation using the Progestin-Primed Ovarian Stimulation (PPOS)"},"summary":{"aims":{"ja":"生殖補助医療において、プロゲスチン製剤（合成プロゲステロン）を排卵抑制剤として用いるProgestin-primed ovarian stimulation（PPOS）法が、調節卵巣刺激法の一つとして普及している。しかし、プロゲスチン製剤が卵胞液、顆粒膜細胞、卵丘細胞などで発現する遺伝子や蛋白に与える影響は十分に評価されていない。本研究はPPOS法で使用されるプロゲスチン製剤が、卵胞発育・卵子の成熟過程に及ぼす影響を、生殖補助医療における採卵及び顕微授精の過程で副次的に得られるヒト卵胞液、卵巣顆粒膜細胞、卵丘細胞、血液検査の残余検体などを用いて従来法と比較検討すること、また、基礎実験の結果や臨床成績を加味して最適なPPOS法を確立すること、を目的としている。","en":"In recent years, the Progestin-Primed Ovarian Stimulation (PPOS) method, utilizing progestin as ovulation suppressants, has gained traction as a controlled ovarian stimulation strategy in assisted reproductive technology. However, the impact of progestin agents on follicular fluid, mural granulosa cell (mGC), and cumulus cells at the levels of gene and protein expression has not been fully assessed. This study aims to evaluate the effects of progestin agents used in the PPOS method, on follicular development, and oocyte maturation."},"methods":{"ja":"層壁顆粒膜細胞（mural granulosa cells：mGC）を採取後洗浄し、Bambanker培地にて凍結保存（-80℃）した。融解後、細胞数を標準化した上でPPOS実施群、従来の排卵誘発法のGnRH-ant実施群、それぞれを混合して解析に用いた。DNase処理後、MidiMACS™ Starting Kit（Miltenyi Biotec社）を用いたソーティング（Dead Cell Removal Kit, Miltenyi Biotec, Bergisch Gladbach, Germany）により死細胞を除去し、Chromium Next GEM Single Cell V(D)J Reagent Kitsを用いて行いてライブラリー調整後、DNBSEQ-G400シーケンサーを用いてsingle cell RNA-seqを実施した。","en":"Collected and washed mGCs were cryopreserved in Bambanker medium at −80 °C. After thawing, cell counts were standardised across patients and normalised to those of the patient with the minimum cell count. Cells were mixed within each group to form the PPOS and GnRH-ant groups. Dead cells were removed through magnetic cell sorting (Dead Cell Removal Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) with LS columns and pre-separation filters from the MidiMACS™ Starting Kit (Miltenyi Biotec), following DNase treatment. After library preparation using Chromium Next GEM Single Cell V(D)J Reagent Kits, single cell RNA-seq was performed using a DNBSEQ-G400 sequencer."},"targets":{"ja":"正常卵巣機能を有する不妊症16症例（PPOS実施群8症例、GnRH-ant実施群 8症例）のmGC","en":"mGCs from 16 infertility patients with normal ovarian function. Eight patients were treated with conventional ovulation induction (GnRH-ant) and eight patients were treated with PPOS."},"url":{"ja":null,"en":null}},"listingSummary":{"methods":{"ja":"発現","en":"Expression profiling"},"targets":{"ja":"正常卵巣機能を有する不妊症：16症例\n（日本人）","en":"16 infertility patients with normal ovarian function\n(Japanese)"},"typeOfData":{"ja":"NGS\n（scRNA-seq）","en":"NGS\n(scRNA-seq)"}},"releaseNote":{"ja":"正常卵巣機能を有する不妊症16症例（PPOS法を実施した8症例、従来の排卵誘発法（GnRH-ant）を実施した8症例）の層壁顆粒膜細胞（mGC）を等量ずつ混和した検体から抽出したRNAを用いたsingle cell RNA-seq解析データをbamファイルにて提供する。","en":"RNAs extracted from granulosa cells which mixed within each group to form the conventional ovulation induction (GnRH-ant) and progestin-primed ovarian stimulation (PPOS) groups were used for single cell RNA-seq analyses. Bam files are provided."},"dataProviders":[{"name":{"ja":"伴田 美佳","en":"Mika Handa"},"organization":{"name":{"ja":"大阪大学大学院医学系研究科 産科学婦人科学講座","en":"Obstetrics and Gynecology, Osaka University"}}}],"researchProjects":[],"grants":[{"title":{"ja":"プロゲスチンが卵子に及ぼす作用機序の解明とPPOS法の最適な治療プロトコルの確立","en":"Elucidating the Mechanism of Progestin Effects on Oocytes and Establishing an Optimal Treatment Protocol for the PPOS Method"},"agency":{"ja":"科学研究費助成事業 基盤研究（C）","en":"KAKENHI Grant-in-Aid for Scientific Research (C)"},"grantIds":["23K08867"]}],"relatedPublications":[{"title":"Adverse effects of progestin-primed ovarian stimulation: Combination of clinical study and single cell analysis","doi":"https://doi.org/10.1016/j.rbmo.2025.104833","datasets":["JGAD000911"]}],"datasets":["JGAD000911"],"controlledAccessUsers":[]}