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成年小鼠背根神经节外植体培养在遗传性神经病变小鼠模型中研究髓鞘形成的优势

Advantages of Adult Mouse Dorsal Root Ganglia Explant Culture in Investigating Myelination in an Inherited Neuropathic Mice Model.

作者信息

Mo Yun Jeong, Kim Yu-Seon, Kim Minseok S, Lee Yun-Il

机构信息

Well Aging Research Center, Division of Biotechnology, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 42988, Korea.

Department of New Biology, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu 42988, Korea.

出版信息

Methods Protoc. 2022 Jul 22;5(4):66. doi: 10.3390/mps5040066.

Abstract

A co-culture of neurons and Schwann cells has frequently been used to investigate myelin sheath formation. However, this approach is restricted to myelin-related diseases of the peripheral nervous system. This study introduces and compares an ex vivo model of adult-mouse-derived dorsal root ganglia (DRG) explant, with an in vitro co-culture of dissociated neurons from mouse embryo DRG and Schwann cells from a mouse sciatic nerve. The 2D co-culture has disadvantages of different mouse isolation for neurons and Schwann cells, animal number, culture duration, and the identification of disease model. However, 3D DRG explant neurons and myelination cells in Matrigel-coated culture are obtained from the same mouse, the culture period is shorter than that of 2D co-culture, and fewer animals are needed. In addition, it has simpler and shorter experimental steps than 2D co-culture. This culture system may prove advantageous in studies of biological functions and pathophysiological mechanisms of disease models, since it can reflect disease characteristics as traditional co-culture does. Therefore, it is suggested that a DRG explant culture is a scientifically, ethically, and economically more practical option than a co-culture system for studying myelin dynamics, myelin sheath formation, and demyelinating disease.

摘要

神经元与雪旺细胞的共培养常被用于研究髓鞘形成。然而,这种方法仅限于外周神经系统的髓鞘相关疾病。本研究引入并比较了一种成年小鼠背根神经节(DRG)外植体的体外模型,以及一种由小鼠胚胎DRG解离神经元和小鼠坐骨神经雪旺细胞组成的体外共培养模型。二维共培养存在神经元和雪旺细胞小鼠分离方式不同、动物数量、培养持续时间以及疾病模型鉴定等缺点。然而,三维DRG外植体神经元和基质胶包被培养中的髓鞘形成细胞取自同一只小鼠,培养周期比二维共培养短,所需动物数量更少。此外,它比二维共培养的实验步骤更简单、更短。这种培养系统在疾病模型的生物学功能和病理生理机制研究中可能具有优势,因为它能像传统共培养一样反映疾病特征。因此,对于研究髓鞘动态、髓鞘形成和脱髓鞘疾病而言,DRG外植体培养被认为是一种在科学、伦理和经济方面比共培养系统更实用的选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d74/9331548/28b207db7c0d/mps-05-00066-g001.jpg

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