从 C9ORF72 相关肌萎缩侧索硬化症患者的外周血单核细胞中生成人类诱导多能干细胞。

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2024-01-01 DOI:10.1007/978-1-0716-3995-5_12
Kalina Andrysiak, Jacek Stępniewski, Magdalena Spaczyńska-Boczar, Katarzyna Łapicka-Bodzioch, Agnieszka Słowik, Józef Dulak
{"title":"从 C9ORF72 相关肌萎缩侧索硬化症患者的外周血单核细胞中生成人类诱导多能干细胞。","authors":"Kalina Andrysiak, Jacek Stępniewski, Magdalena Spaczyńska-Boczar, Katarzyna Łapicka-Bodzioch, Agnieszka Słowik, Józef Dulak","doi":"10.1007/978-1-0716-3995-5_12","DOIUrl":null,"url":null,"abstract":"<p><p>Disease modeling of neuromuscular disorders, such as amyotrophic lateral sclerosis (ALS), is hindered by limited accessibility of affected cells. This problem can be overcome by generation of human induced pluripotent stem cells (hiPSC), which can be then differentiated into required cells. Here, we describe the detailed protocol of hiPSC establishment from peripheral blood mononuclear cells (PBMC) of two ALS patients with detected expansion of G4C2 (GGGGCC) repeats in the first intron of C9ORF72 gene, known to be linked with the most common form of familial ALS.Successful PBMC reprogramming with non-integrating Sendai vectors was confirmed by expression of pluripotency markers: OCT4, NANOG, SSEA4, and TRA-1-60 in obtained hiPSC and their ability to differentiate into cells of three germ layers.The generated ALS-patient-specific hiPSC create a possibility for deciphering molecular basis of this devastating neuromuscular disease.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2835 ","pages":"135-146"},"PeriodicalIF":0.0000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Generation of Human-Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells of C9ORF72-Associated Amyotrophic Lateral Sclerosis Patients.\",\"authors\":\"Kalina Andrysiak, Jacek Stępniewski, Magdalena Spaczyńska-Boczar, Katarzyna Łapicka-Bodzioch, Agnieszka Słowik, Józef Dulak\",\"doi\":\"10.1007/978-1-0716-3995-5_12\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Disease modeling of neuromuscular disorders, such as amyotrophic lateral sclerosis (ALS), is hindered by limited accessibility of affected cells. This problem can be overcome by generation of human induced pluripotent stem cells (hiPSC), which can be then differentiated into required cells. Here, we describe the detailed protocol of hiPSC establishment from peripheral blood mononuclear cells (PBMC) of two ALS patients with detected expansion of G4C2 (GGGGCC) repeats in the first intron of C9ORF72 gene, known to be linked with the most common form of familial ALS.Successful PBMC reprogramming with non-integrating Sendai vectors was confirmed by expression of pluripotency markers: OCT4, NANOG, SSEA4, and TRA-1-60 in obtained hiPSC and their ability to differentiate into cells of three germ layers.The generated ALS-patient-specific hiPSC create a possibility for deciphering molecular basis of this devastating neuromuscular disease.</p>\",\"PeriodicalId\":18490,\"journal\":{\"name\":\"Methods in molecular biology\",\"volume\":\"2835 \",\"pages\":\"135-146\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Methods in molecular biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1007/978-1-0716-3995-5_12\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/978-1-0716-3995-5_12","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0

摘要

神经肌肉疾病(如肌萎缩性脊髓侧索硬化症(ALS))的疾病建模受到受影响细胞获取途径有限的阻碍。这个问题可以通过生成人类诱导多能干细胞(hiPSC)来解决,这种干细胞可以分化成所需的细胞。在这里,我们描述了从两名 ALS 患者的外周血单核细胞(PBMC)中建立 hiPSC 的详细方案,这两名 ALS 患者的 C9ORF72 基因第一个内含子中检测到 G4C2 (GGGGCC) 重复序列扩增,已知与最常见的家族性 ALS 有关:用非整合仙台病毒载体对 PBMC 进行重编程的成功,通过获得的 hiPSC 中 OCT4、NANOG、SSEA4 和 TRA-1-60 等多能性标记的表达及其分化为三个胚层细胞的能力得到了证实。
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
Generation of Human-Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells of C9ORF72-Associated Amyotrophic Lateral Sclerosis Patients.

Disease modeling of neuromuscular disorders, such as amyotrophic lateral sclerosis (ALS), is hindered by limited accessibility of affected cells. This problem can be overcome by generation of human induced pluripotent stem cells (hiPSC), which can be then differentiated into required cells. Here, we describe the detailed protocol of hiPSC establishment from peripheral blood mononuclear cells (PBMC) of two ALS patients with detected expansion of G4C2 (GGGGCC) repeats in the first intron of C9ORF72 gene, known to be linked with the most common form of familial ALS.Successful PBMC reprogramming with non-integrating Sendai vectors was confirmed by expression of pluripotency markers: OCT4, NANOG, SSEA4, and TRA-1-60 in obtained hiPSC and their ability to differentiate into cells of three germ layers.The generated ALS-patient-specific hiPSC create a possibility for deciphering molecular basis of this devastating neuromuscular disease.

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
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
期刊最新文献
A Guideline Strategy for Identifying a Viral Gene/Protein Evading Antiviral Innate Immunity. A Guideline Strategy for Identifying Genes/Proteins Regulating Antiviral Innate Immunity. Application of Proteomics Technology Based on LC-MS Combined with Western Blotting and Co-IP in Antiviral Innate Immunity. Click Chemistry in Detecting Protein Modification. CRISPR-Mediated Construction of Gene-Knockout Mice for Investigating Antiviral Innate Immunity.
×
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