Landegger Lukas D, Dilwali Sonam, Stankovic Konstantina M
Eaton Peabody Laboratories, Department of Otolaryngology, Massachusetts Eye and Ear; Department of Otolaryngology, Harvard Medical School; Department of Otolaryngology, Vienna General Hospital, Medical University of Vienna.
Eaton Peabody Laboratories, Department of Otolaryngology, Massachusetts Eye and Ear; Harvard Program in Speech and Hearing Bioscience and Technology.
J Vis Exp. 2017 Jun 8(124):55704. doi: 10.3791/55704.
While there have been remarkable advances in hearing research over the past few decades, there is still no cure for Sensorineural Hearing Loss (SNHL), a condition that typically involves damage to or loss of the delicate mechanosensory structures of the inner ear. Sophisticated in vitro and ex vivo assays have emerged in recent years, enabling the screening of an increasing number of potentially therapeutic compounds while minimizing resources and accelerating efforts to develop cures for SNHL. Though homogenous cultures of certain cell types continue to play an important role in current research, many scientists now rely on more complex organotypic cultures of murine inner ears, also known as cochlear explants. The preservation of organized cellular structures within the inner ear facilitates the in situ evaluation of various components of the cochlear infrastructure, including inner and outer hair cells, spiral ganglion neurons, neurites, and supporting cells. Here we present the preparation, culture, treatment, and immunostaining of neonatal murine cochlear explants. The careful preparation of these explants facilitates the identification of mechanisms that contribute to SNHL and constitutes a valuable tool for the hearing research community.
尽管在过去几十年里听力研究取得了显著进展,但感音神经性听力损失(SNHL)仍然无法治愈,这种病症通常涉及内耳精细的机械感觉结构受损或丧失。近年来出现了复杂的体外和离体检测方法,能够在减少资源消耗的同时筛选越来越多潜在的治疗化合物,并加快开发SNHL治疗方法的努力。虽然某些细胞类型的同质培养物在当前研究中仍然发挥着重要作用,但现在许多科学家依赖于更复杂的小鼠内耳器官型培养物,也称为耳蜗外植体。内耳内有组织的细胞结构的保存有助于对耳蜗基础设施的各种组成部分进行原位评估,包括内、外毛细胞、螺旋神经节神经元、神经突和支持细胞。在这里,我们介绍新生小鼠耳蜗外植体的制备、培养、处理和免疫染色。这些外植体的精心制备有助于确定导致SNHL的机制,是听力研究界的一个有价值的工具。