幼鼠内耳单个器官培养步骤及分类方法

Technical Details of Sample Preparation for Inner Ear Organotypic Cultures

内耳离体培养在研究内耳细胞功能、内耳组织发育再生、细胞损伤退化等方面具有重要价值。但由于内耳的组织结构微小复杂,其取材和培养具有较大难度,使该技术的普及和应用受到一定限制。本文在简要点评不同内耳离体培养方法优劣的基础上,结合本实验室多年来积累的实践经验,详细介绍了新生大鼠各个内耳终器取材及培养的具体步骤。从解剖迷路到分离终器,从凝胶制备到标本铺放,都有详细的操作指南,本文还针对各环节中的技术动作和关键事项进行了具体探讨并配以指导图例,希望这些内容能够弥补以往文献中抽象简略的条款式步骤所造成的实验困难,为开展内耳器官培养研究的初学者提供一定的帮助。

In vitro culture techniques have important values in the investigation of inner ear cellular functions, organic development, cell degeneration and/or cell regeneration. Since structures of the inner ear are tiny and complex, sample prepara- tion for organotypic culture of the inner ear can be difficult, despite that relevant techniques have been introduced briefly in numerous earlier publications. Based upon decades of culturing practices with inner ear end-organs in this lab, this article ad- dresses the technical aspects of inner ear organic culturing by firstly presenting a brief review of various culture methods for different inner ear materials. Detailed step by step protocols for sample preparation, collection and culture are also described. This presentation provides detailed operative guidelines regarding useful techniques and tricks in micro-dissection of the whole membranous labyrinth, in sample preparation, collagen gel preparation and sample layer mountings with detailed illus- trations. We hope this article can serve as an additional reading material supplementing previous literatures that lack detailed technical information for beginners in this field.