{"id":142,"title":"A Three-Dimensional Gelatin Methacryloyl (GelMA) Hydrogel Platform for In Vitro Bovine Ovarian Tissue Culture and Primordial Follicle Activation","title_zh":"三维GelMA水凝胶平台用于牛卵巢组织体外培养与原始卵泡激活","url":"https://pubmed.ncbi.nlm.nih.gov/42992674","doi":"10.3791/73725","pmid":"42992674","journal":"J Vis Exp","kind":"paper","lang":"en","published_at":"2026-11-05T00:00:00+00:00","fetched_at":"2026-11-08T03:43:57+00:00","authors":["Liu M","Xu M","Wang S","Wu W","Yau KMY","Xiang Y","Tsui MTK","Poon LC","Chan DYL"],"primary_topic":"oocyte","topics":[{"slug":"oocyte","score":13.0,"hits":["follicle","follicular","folliculogenesis","granulosa","oocyte","ovarian","primordial follicle"]},{"slug":"fertility-preservation","score":4.0,"hits":["ovarian tissue"]}],"topic_score":13.0,"preference_score":0.0,"preference_labels":[],"llm_score":72.0,"final_score":69.0,"score_dims":{"novelty":62,"rigor":72,"impact":65,"relevance":82,"translation":70},"recommend_reason":"方法学文章，把可光交联的GelMA三维培养方案标准化，并给出与2D、Matrigel的对照结果；对做卵巢组织培养、卵泡激活与生育力保存的实验室有直接可操作价值，但所用模型为牛，机制新意有限。","keywords":["GelMA水凝胶","三维培养","卵巢组织","原始卵泡激活","牛模型","生育力保存"],"selected":1,"story_id":58,"llm_provider":"deepseek","llm_model":"deepseek-flash","llm_effort":"low","source_key":"pubmed-gametes","source_name":"PubMed · 配子与性腺","source_tier":1,"mention_count":1,"has_interpretation":false,"journal_key":"jove","journal_tier":"low-signal","journal_if":1.2,"is_mock":false,"summary_zh":"该研究方案采用4%(w/v)甲基丙烯酰化明胶（GelMA）水凝胶支架，以0.1%(w/v)LAP为光引发剂，将1 mm³牛卵巢皮质碎片包埋后光交联，在38.5 ℃、5% CO₂条件下体外培养6天。研究通过中性红染色检测培养前卵泡活力，并用H&E染色、扫描电镜观察支架结构，同时定量分析卵泡直径与发育阶段分布；结果显示，与常规模拟（2D）培养及Matrigel体系相比，该平台能更好地维持三维卵泡结构，并促进原始卵泡激活及向次级卵泡阶段进展。该方法所用材料均为市售，重复性好，为大动物模型中的早期卵泡发生研究提供了生理相关性较强的平台。","summary":"Conventional two-dimensional (2D) culture systems fail to preserve the three-dimensional (3D) follicular architecture, permitting granulosa cell migration away from the oocyte and disrupting paracrine signaling essential for folliculogenesis. Hydrogel-based 3D culture platforms have emerged as a promising alternative for maintaining tissue integrity during extended in vitro culture. This protocol describes a 3D ovarian tissue culture method using 4% (w/v) gelatin methacryloyl (GelMA) hydrogel scaffolds with 0.1% (w/v) lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) as the photoinitiator. Bovine ovarian cortical fragments (1 mm × 1 mm × 1 mm) embedded in photocrosslinked GelMA scaffolds are cultured for 6 days under standard conditions (38.5 °C, 5% CO₂). Compared with conventional 2D culture and Matrigel-based systems, this method enhances primordial follicle activation and progression to the secondary stage, as confirmed by follicle stage distribution analysis and diameter measurements. Key outcome measures include baseline follicle viability assessed by neutral red staining prior to culture, histological analysis with hematoxylin and eosin (H&E) staining, scanning electron microscopy (SEM) of scaffold architecture, and quantitative assessment of follicle diameter and developmental stage distribution. The protocol is reproducible, relies on commercially available materials, and provides a physiologically relevant platform for investigating primordial follicle activation and early folliculogenesis in large animal models.","interpretation_basis":null,"fulltext_status":null,"interpretation":null,"fulltext":{"available":false,"status":null,"reason":"尚未抓取全文（未入选、分数未达阈值，或还没跑 fulltext 步骤）","pmcid":null,"license":null,"chars":0,"source":null,"fetched_at":null}}