The Effects of Co-Culturing ND7/23 Sensory Neuron-like Cells and IFRS1 Schwann Cells on Myelination: A Single-Arm Nonrandomized Study.

IF 3 Q2 CLINICAL NEUROLOGY
Shizuka Takaku, Kazunori Sango
{"title":"The Effects of Co-Culturing ND7/23 Sensory Neuron-like Cells and IFRS1 Schwann Cells on Myelination: A Single-Arm Nonrandomized Study.","authors":"Shizuka Takaku, Kazunori Sango","doi":"10.3390/neurolint17090138","DOIUrl":null,"url":null,"abstract":"<p><p><b>Background/Objectives</b>: Co-culture models of neurons and Schwann cells have been used to explore the mechanisms of myelination during development, axonal regeneration after injury, and the pathogenesis of various demyelinating neuropathies. A spontaneously immortalized Fischer rat Schwann cell line 1 (IFRS1), established from the primary culture of adult Fischer344 rat peripheral nerves, can myelinate neurites in co-cultures with primary cultured dorsal root ganglion neurons and neuronal cell lines, such as nerve growth factor (NGF)-primed PC12 cells and NSC-34 motor neuron-like cells. In this study, we aimed to establish a stable co-culture system using IFRS1 cells and ND7/23 sensory neuron-like cells. <b>Methods</b>: ND7/23 cells were seeded at a low density (2 × 10<sup>3</sup>/cm<sup>2</sup>) and maintained for 7 days in serum-containing medium supplemented with NGF (10 ng/mL) and the Rho kinase inhibitor Y27632 (5 μM) to promote neurite elongation. The cells were then treated with the anti-mitotic agent mitomycin C (1 μg/mL) for 12-16 h to suppress proliferative activity. Following this, the cells were co-cultured with IFRS1 cells (2 × 10<sup>4</sup>/cm<sup>2</sup>) and maintained at 37 °C in serum-containing medium supplemented with ascorbic acid (50 μg/mL), NGF (10 ng/mL), and ciliary neurotrophic factor (10 ng/mL). <b>Results</b>: Double-immunofluorescence staining performed on day 21 of the co-culture revealed myelin protein 22- or myelin basic protein-immunoreactive IFRS1 cells surrounding βIII tubulin-immunoreactive neurites emerging from ND7/23 cells. Myelin formation was further confirmed via Sudan Black B staining and electron microscopy. <b>Conclusions</b>: This co-culture system may provide a valuable tool for studying the processes of myelination in the peripheral nervous system, as well as the pathogenesis of various sensory neuropathies and potential novel therapeutic approaches for these conditions.</p>","PeriodicalId":19130,"journal":{"name":"Neurology International","volume":"17 9","pages":""},"PeriodicalIF":3.0000,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472592/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Neurology International","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3390/neurolint17090138","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CLINICAL NEUROLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Background/Objectives: Co-culture models of neurons and Schwann cells have been used to explore the mechanisms of myelination during development, axonal regeneration after injury, and the pathogenesis of various demyelinating neuropathies. A spontaneously immortalized Fischer rat Schwann cell line 1 (IFRS1), established from the primary culture of adult Fischer344 rat peripheral nerves, can myelinate neurites in co-cultures with primary cultured dorsal root ganglion neurons and neuronal cell lines, such as nerve growth factor (NGF)-primed PC12 cells and NSC-34 motor neuron-like cells. In this study, we aimed to establish a stable co-culture system using IFRS1 cells and ND7/23 sensory neuron-like cells. Methods: ND7/23 cells were seeded at a low density (2 × 103/cm2) and maintained for 7 days in serum-containing medium supplemented with NGF (10 ng/mL) and the Rho kinase inhibitor Y27632 (5 μM) to promote neurite elongation. The cells were then treated with the anti-mitotic agent mitomycin C (1 μg/mL) for 12-16 h to suppress proliferative activity. Following this, the cells were co-cultured with IFRS1 cells (2 × 104/cm2) and maintained at 37 °C in serum-containing medium supplemented with ascorbic acid (50 μg/mL), NGF (10 ng/mL), and ciliary neurotrophic factor (10 ng/mL). Results: Double-immunofluorescence staining performed on day 21 of the co-culture revealed myelin protein 22- or myelin basic protein-immunoreactive IFRS1 cells surrounding βIII tubulin-immunoreactive neurites emerging from ND7/23 cells. Myelin formation was further confirmed via Sudan Black B staining and electron microscopy. Conclusions: This co-culture system may provide a valuable tool for studying the processes of myelination in the peripheral nervous system, as well as the pathogenesis of various sensory neuropathies and potential novel therapeutic approaches for these conditions.

ND7/23感觉神经元样细胞和IFRS1雪旺细胞共培养对髓鞘形成的影响:一项单组非随机研究
背景/目的:利用神经元与雪旺细胞共培养模型,探讨发育过程中髓鞘形成的机制、损伤后轴突的再生以及各种脱髓鞘神经病变的发病机制。以成年Fischer344大鼠周围神经原代培养为基础,建立了一种自发永活的Fischer大鼠Schwann细胞系1 (IFRS1),可与原代培养的背根神经节神经元和神经生长因子(NGF)诱导的PC12细胞和NSC-34运动神经元样细胞等神经细胞系共培养,形成髓鞘神经突。本研究旨在利用IFRS1细胞和ND7/23感觉神经元样细胞建立稳定的共培养体系。方法:将ND7/23细胞低密度(2 × 103/cm2)接种于添加NGF (10 ng/mL)和Rho激酶抑制剂Y27632 (5 μM)的含血清培养基中,培养7 d,促进神经突延长。用抗有丝分裂剂丝裂霉素C (1 μg/mL)作用细胞12 ~ 16 h,抑制细胞增殖活性。随后,将细胞与IFRS1细胞(2 × 104/cm2)共培养,在含抗坏血酸(50 μg/mL)、NGF (10 ng/mL)和纤毛神经营养因子(10 ng/mL)的含血清培养基中37℃保存。结果:共培养第21天,双免疫荧光染色显示ND7/23细胞中出现βIII微管蛋白免疫反应神经突周围的髓鞘蛋白22或髓鞘碱性蛋白免疫反应IFRS1细胞。通过苏丹黑B染色和电镜进一步证实髓磷脂的形成。结论:该共培养系统可能为研究周围神经系统髓鞘形成过程、各种感觉神经病变的发病机制和潜在的新治疗方法提供有价值的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Neurology International
Neurology International CLINICAL NEUROLOGY-
CiteScore
3.70
自引率
3.30%
发文量
69
审稿时长
11 weeks
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信