Separation of Avian Preovulatory Follicle Granulosa and Theca Cell Layers for Downstream Applications.

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
Ashley E Kramer, Kathryn M Ellwood, Erin M Brannick, Aditya Dutta
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引用次数: 0

Abstract

Layer hens (egg-laying chickens) and broiler breeders (breeding stock for meat-producing chickens) are crucial to the world's food supply as a reliable source of protein. They are also an emerging animal model for the study of human reproductive disease. As the field of poultry research develops, the health and function of the layer hen and broiler breeder ovary will be an important point of study for both agricultural and biomedical researchers. One of the challenges presented by this emerging interest is the need for replicable techniques that all researchers can employ in ovarian specimen collection. In particular, a detailed visual process must be established to define the proper separation of the specialized granulosa and theca cell layers from hen follicles to achieve agreement and consistency among researchers. This study describes the extraction of preovulatory follicles and ovary tissue in white leghorn hens of prime reproductive age. The separation of these follicles is performed under cold, liquid conditions to congeal the yolk for easier manipulation and to prevent the follicle's own weight from tearing apart cell layers during the separation process. Once the separation is complete, the desired cell layers can be further digested for tissue culture approaches or can be cryopreserved for genomic and proteomic analyses.

为下游应用分离禽类排卵前卵泡颗粒细胞和子宫颈细胞层。
蛋鸡(产蛋鸡)和肉用种鸡(产肉鸡的种群)作为可靠的蛋白质来源,对世界粮食供应至关重要。它们也是研究人类生殖疾病的新兴动物模型。随着家禽研究领域的发展,蛋鸡和肉用种鸡卵巢的健康和功能将成为农业和生物医学研究人员的重要研究对象。这一新兴兴趣所带来的挑战之一是需要所有研究人员都能在卵巢标本采集中使用的可复制技术。特别是,必须建立详细的可视化流程,以确定如何从母鸡卵泡中正确分离特化的颗粒细胞层和卵巢细胞层,从而实现研究人员之间的一致性。本研究介绍了如何提取处于生育黄金期的白羽蛋鸡的排卵前卵泡和卵巢组织。卵泡的分离是在低温、液态条件下进行的,目的是使卵黄凝固,便于操作,并防止卵泡自身重量在分离过程中撕裂细胞层。分离完成后,所需的细胞层可进一步消化以进行组织培养,或低温保存以进行基因组和蛋白质组分析。
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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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