Basic fibroblast growth factor promotes the development of human ovarian early follicles during growth in vitro
Basic fibroblast growth factor promotes the development of human ovarian early follicles during growth in vitro
复制标题
碱性成纤维细胞生长因子促进人卵巢早期卵泡体外生长过程中的发育
DOI：
10.1093/humrep/det465
复制
发表时间：
2014-03-01
影响因子：
6.1
通讯作者：
Qiao, Jie
中科院分区：
文献类型：
--
作者：
Wang, Tian-ren;Yan, Li-ying;Qiao, Jie
研究问题：在体外三维培养体系中，碱性成纤维细胞生长因子对人早期卵泡生长有何影响？总结回答：添加200 ng/ml的碱性成纤维细胞生长因子可促进人早期卵泡的生长、存活和体外生长过程中的存活率。然而，个体早期卵泡在囊化3D培养中的生长和存活还没有报道。研究设计、大小、持续时间：研究了人卵巢卵泡的体外成熟。卵巢组织11例，取自11例妇科疾病腹腔镜手术患者，均征得书面知情同意。用酶消化法和机械破碎法分离出154个早期卵泡。将它们分别包裹在1%w/v的藻酸盐中，随机分为0、100、200和300 ng/ml的碱性成纤维细胞生长因子培养8天。显微镜下观察卵泡存活和生长情况。在激光共聚焦显微镜下观察卵泡的存活率，并进行钙黄素-AM和同型二聚体-I(Ca-AM/ethD-I)染色。MAIN结果和随机因素的作用：培养8天后，154个卵泡全部增大。含200 ng bFGF/ml组的卵泡直径、存活率及存活率均显著高于未加bFGF组(P<；0.05)。200 ng bFGF/ml组的腔前卵泡百分率明显高于不加bFGF组(P<；0.05)，而原始卵泡和初级卵泡的百分率则显著低于对照组(P<；0.05)。限制，谨慎的理由：本研究主要集中在体外3D培养中对单个人早期卵泡发育的影响，但对揭示不同阶段bFGF的具体作用能力有限。这一发现突显了改进人卵巢卵泡获取和分离的必要性。研究结果的广泛意义：在适当剂量的碱性成纤维细胞生长因子的作用下，人卵泡的体外3D培养为研究其发育提供了一种有效的方法。此外，它可以将早期卵泡培养到高级阶段，因此有可能成为辅助生殖技术的成熟卵母细胞的重要来源；特别是作为保留妇女生育能力的一种选择，包括癌症患者。
STUDY QUESTION: What is the effect of basic fibroblast growth factor (bFGF) on the growth of individual early human follicles in a three-dimensional (3D) culture system in vitro?SUMMARY ANSWER: The addition of 200 ng bFGF/ml improves human early follicle growth, survival and viability during growth in vitro.WHAT IS KNOWN ALREADY: It has been demonstrated that bFGF enhances primordial follicle development in human ovarian tissue culture. However, the growth and survival of individual early follicles in encapsulated 3D culture have not been reported.STUDY DESIGN, SIZE, DURATION: The maturation in vitro of human ovarian follicles was investigated. Ovarian tissue (n = 11) was obtained from 11 women during laparoscopic surgery for gynecological disease, after obtaining written informed consent. One hundred and fifty-four early follicles were isolated by enzymic digestion and mechanical disruption. They were individually encapsulated into alginate (1% w/v) and randomly assigned to be cultured with 0, 100, 200 or 300 ng bFGF/ml for 8 days.PARTICIPANTS/MATERIALS, SETTING, METHODS: Individual follicles were cultured in minimum essential medium alpha (alpha MEM) supplemented with bFGF. Follicle survival and growth were assessed by microscopy. Follicle viability was evaluated under confocal laser scanning microscope following Calcein-AM and Ethidium homodimer-I (Ca-AM/EthD-I) staining.MAIN RESULTS AND THE ROLE OF CHANCE: After 8 days in culture, all 154 follicles had increased in size. The diameter and survival rate of the follicles and the percentage with good viability were significantly higher in the group cultured with 200 ng bFGF/ml than in the group without bFGF (P < 0.05). The percentage of follicles in the pre-antral stage was significantly higher in the 200 ng bFGF/ml group than in the group without bFGF (P < 0.05), while the percentages of primordial and primary follicles were significantly lower (P < 0.05).LIMITATIONS, REASONS FOR CAUTION: The study focuses on the effect of bFGF on the development of individual human early follicles in 3D culture in vitro and has limited ability to reveal the specific effect of bFGF at each different stage. The findings highlight the need to improve the acquisition and isolation of human ovarian follicles.WIDER IMPLICATIONS OF THE FINDINGS: The in vitro 3D culture of human follicles with appropriate dosage of bFGF offers an effective method to investigate their development. Moreover, it allows early follicles to be cultured to an advanced stage and therefore has the potential to become an important source of mature oocytes for assisted reproductive technology; particularly as an option for fertility preservation in women, including patients with cancer.