大鼠幼鼠嗅粘膜上皮细胞的纯度测定

Joghataei Mohammad Taghi, Bakhtiari Mehrdad, F. Farhid, Z. Mojgan, I. Mohammad, Moaieri Ardshir, Azizi Monireh
{"title":"大鼠幼鼠嗅粘膜上皮细胞的纯度测定","authors":"Joghataei Mohammad Taghi, Bakhtiari Mehrdad, F. Farhid, Z. Mojgan, I. Mohammad, Moaieri Ardshir, Azizi Monireh","doi":"10.18869/ACADPUB.JBRMS.3.4.12","DOIUrl":null,"url":null,"abstract":"Introduction: Cell transplantation is one of the main strategies for spinal cord injury repair. As OECs of the olfactory mucosa can be obtained by simple biopsy in all individuals without affecting their smell sensation, OECs considered as a promising candidate for autologous transplantation in the nervous system injury, especially for spinal cord repair. Thereby in the current study OECs were cultured from olfactory mucosa of 7 days old rats' pups and their purity was examined by flow-cytometry after simultaneous double staining for p75 and GFAP markers. Materials and methods: 7 days old Wistar rats' pups were deeply anesthetized by ketamine / xylazine (60/6mg/Kg). Then the nasal cavity was opened sagittally and the olfactory mucosa was separated from posterior part of nasal septum and at last OECs were obtained from lamina properia of olfactory mucosa and were cultured. The cultured cells were simultaneously immunolabeled for p75 and GFAP markers and finally purity of cultured cells assessed by flow-cytometry. Results: cultured OECs demonstrated two different morphologies, a spindle shape Schwannlike and an astrocyte-like OECs with flat sheet-like morphology. In addition, simultaneous immunolabeling for p75 and GFAP markers of OECs exhibited OECs were positive for both markers at the same time. The flow-cytometry results displayed that 87.9±2.4% of cells were p75/ GFAP double positive cells,1.05±0.4 only p75 positive and 5.8±1.5% were single positive for GFAP. Conclusion: Purity of cultured OECs in our study is probably more than 87.9% by flowowing to p75+/S100+ and GFAP+/S100+ olfactory ensheathing cells were not counted. Thus the culture procedure of this study seems to be a good protocol for OECs purifying and cell therapy in CNS damages.","PeriodicalId":15047,"journal":{"name":"Journal of Basic Research in Medical Sciences","volume":"3 1","pages":"12-21"},"PeriodicalIF":0.0000,"publicationDate":"2016-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"7","resultStr":"{\"title\":\"Purity determining of cultured OECs from olfactory mucosa of rats' pups\",\"authors\":\"Joghataei Mohammad Taghi, Bakhtiari Mehrdad, F. Farhid, Z. Mojgan, I. Mohammad, Moaieri Ardshir, Azizi Monireh\",\"doi\":\"10.18869/ACADPUB.JBRMS.3.4.12\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Introduction: Cell transplantation is one of the main strategies for spinal cord injury repair. As OECs of the olfactory mucosa can be obtained by simple biopsy in all individuals without affecting their smell sensation, OECs considered as a promising candidate for autologous transplantation in the nervous system injury, especially for spinal cord repair. Thereby in the current study OECs were cultured from olfactory mucosa of 7 days old rats' pups and their purity was examined by flow-cytometry after simultaneous double staining for p75 and GFAP markers. Materials and methods: 7 days old Wistar rats' pups were deeply anesthetized by ketamine / xylazine (60/6mg/Kg). Then the nasal cavity was opened sagittally and the olfactory mucosa was separated from posterior part of nasal septum and at last OECs were obtained from lamina properia of olfactory mucosa and were cultured. The cultured cells were simultaneously immunolabeled for p75 and GFAP markers and finally purity of cultured cells assessed by flow-cytometry. Results: cultured OECs demonstrated two different morphologies, a spindle shape Schwannlike and an astrocyte-like OECs with flat sheet-like morphology. In addition, simultaneous immunolabeling for p75 and GFAP markers of OECs exhibited OECs were positive for both markers at the same time. The flow-cytometry results displayed that 87.9±2.4% of cells were p75/ GFAP double positive cells,1.05±0.4 only p75 positive and 5.8±1.5% were single positive for GFAP. Conclusion: Purity of cultured OECs in our study is probably more than 87.9% by flowowing to p75+/S100+ and GFAP+/S100+ olfactory ensheathing cells were not counted. Thus the culture procedure of this study seems to be a good protocol for OECs purifying and cell therapy in CNS damages.\",\"PeriodicalId\":15047,\"journal\":{\"name\":\"Journal of Basic Research in Medical Sciences\",\"volume\":\"3 1\",\"pages\":\"12-21\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Basic Research in Medical Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.18869/ACADPUB.JBRMS.3.4.12\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Basic Research in Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.18869/ACADPUB.JBRMS.3.4.12","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 7

摘要

细胞移植是脊髓损伤修复的主要策略之一。由于嗅觉粘膜的oec可以在所有个体中通过简单的活检获得,而不影响其嗅觉感觉,因此oec被认为是神经系统损伤,特别是脊髓修复中自体移植的有希望的候选者。因此,本研究从7日龄大鼠幼鼠嗅粘膜中培养oec,并通过流式细胞术同时对p75和GFAP标记物进行双重染色,检测oec的纯度。材料与方法:采用氯胺酮/噻嗪(60/6mg/Kg)深度麻醉7日龄Wistar大鼠幼鼠。然后矢状打开鼻腔,从鼻中隔后部分离嗅觉黏膜,最后从嗅觉黏膜固有层获得oec并进行培养。同时对培养细胞进行p75和GFAP标记免疫标记,最后用流式细胞术评估培养细胞的纯度。结果:培养的oec表现出两种不同的形态,一种是纺锤形雪旺样oec,另一种是星形细胞样oec,具有扁平片状形态。此外,oec的p75和GFAP标记同时免疫标记显示oec对这两种标记同时呈阳性。流式细胞术结果显示,87.9±2.4%的细胞为p75/ GFAP双阳性细胞,1.05±0.4的细胞为p75阳性细胞,5.8±1.5%的细胞为GFAP单阳性细胞。结论:经p75+/S100+和GFAP+/S100+嗅鞘细胞流动,本研究培养的嗅鞘细胞纯度可达87.9%以上。因此,本研究的培养过程似乎是一种良好的oec纯化和细胞治疗中枢神经系统损伤的方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Purity determining of cultured OECs from olfactory mucosa of rats' pups
Introduction: Cell transplantation is one of the main strategies for spinal cord injury repair. As OECs of the olfactory mucosa can be obtained by simple biopsy in all individuals without affecting their smell sensation, OECs considered as a promising candidate for autologous transplantation in the nervous system injury, especially for spinal cord repair. Thereby in the current study OECs were cultured from olfactory mucosa of 7 days old rats' pups and their purity was examined by flow-cytometry after simultaneous double staining for p75 and GFAP markers. Materials and methods: 7 days old Wistar rats' pups were deeply anesthetized by ketamine / xylazine (60/6mg/Kg). Then the nasal cavity was opened sagittally and the olfactory mucosa was separated from posterior part of nasal septum and at last OECs were obtained from lamina properia of olfactory mucosa and were cultured. The cultured cells were simultaneously immunolabeled for p75 and GFAP markers and finally purity of cultured cells assessed by flow-cytometry. Results: cultured OECs demonstrated two different morphologies, a spindle shape Schwannlike and an astrocyte-like OECs with flat sheet-like morphology. In addition, simultaneous immunolabeling for p75 and GFAP markers of OECs exhibited OECs were positive for both markers at the same time. The flow-cytometry results displayed that 87.9±2.4% of cells were p75/ GFAP double positive cells,1.05±0.4 only p75 positive and 5.8±1.5% were single positive for GFAP. Conclusion: Purity of cultured OECs in our study is probably more than 87.9% by flowowing to p75+/S100+ and GFAP+/S100+ olfactory ensheathing cells were not counted. Thus the culture procedure of this study seems to be a good protocol for OECs purifying and cell therapy in CNS damages.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
审稿时长
6 weeks
期刊最新文献
Investigating the factors and methods of suicide in Ilam province (from 2012 to 2016) Fungal infection in foot diabetic patients Ureteral stone extraction under direct ureteroscopic vision: An alternative for routine ureteral stone management Association of low serum level of secreted frizzled-related protein 5 (SFRP5) with the presence and severity of coronary artery disease Lack of association between insulin-like growth factor-I (IGF-I) gene polymorphism (rs35767) and power performance in professional, amateur athletes and non-athlete individuals in Iranian population
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1