人脐带血间充质干细胞/祖细胞的渗透选择。

Biju Parekkadan, Palaniappan Sethu, Daan van Poll, Martin L Yarmush, Mehmet Toner
{"title":"人脐带血间充质干细胞/祖细胞的渗透选择。","authors":"Biju Parekkadan,&nbsp;Palaniappan Sethu,&nbsp;Daan van Poll,&nbsp;Martin L Yarmush,&nbsp;Mehmet Toner","doi":"10.1089/ten.2007.0054","DOIUrl":null,"url":null,"abstract":"<p><p>The isolation of undifferentiated adult stem/progenitor cells remains a challenging task primarily due to the rare quantity of these cells in biological samples and the lack of unique markers. Herein, we report a relatively straightforward method for isolation of human mesenchymal stem cells (MSCs) based on their unusual resistance to osmotic lysis, which we term \"osmotic selection\" (OS). MSCs can remarkably withstand significant exposure to hypotonic conditions (> 30 min) with only a reversible impairment in cell proliferation and with no loss of stem cell potential after exposure. Comparison of MSCs to other circulating nonhematopoietic cells revealed a time regime, by which purification of these cells would be attainable without considerable cell loss. OS showed a 50-fold enrichment of fibroblast colony-forming units from umbilical cord blood samples when compared to commonly employed techniques. After upstream processing, isolated cells using OS were immunophenotyped to be CD14-, CD34-, CD45-, CD44+, CD105+, and CD106+, and displayed multipotent differentiation. Preliminary investigations to determine mechanisms responsible for osmolytic resistance revealed MSCs to have an ineffective volume of 59%, with the ability to double cell volume at infinite dilution. Disruption of filamentous actin polymerization by cytochalasin D sensitized MSCs to osmotic lysis, which suggests a cytoskeletal element involved in osmolytic resistance.</p>","PeriodicalId":23102,"journal":{"name":"Tissue engineering","volume":"13 10","pages":"2465-73"},"PeriodicalIF":0.0000,"publicationDate":"2007-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/ten.2007.0054","citationCount":"28","resultStr":"{\"title\":\"Osmotic selection of human mesenchymal stem/progenitor cells from umbilical cord blood.\",\"authors\":\"Biju Parekkadan,&nbsp;Palaniappan Sethu,&nbsp;Daan van Poll,&nbsp;Martin L Yarmush,&nbsp;Mehmet Toner\",\"doi\":\"10.1089/ten.2007.0054\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The isolation of undifferentiated adult stem/progenitor cells remains a challenging task primarily due to the rare quantity of these cells in biological samples and the lack of unique markers. Herein, we report a relatively straightforward method for isolation of human mesenchymal stem cells (MSCs) based on their unusual resistance to osmotic lysis, which we term \\\"osmotic selection\\\" (OS). MSCs can remarkably withstand significant exposure to hypotonic conditions (> 30 min) with only a reversible impairment in cell proliferation and with no loss of stem cell potential after exposure. Comparison of MSCs to other circulating nonhematopoietic cells revealed a time regime, by which purification of these cells would be attainable without considerable cell loss. OS showed a 50-fold enrichment of fibroblast colony-forming units from umbilical cord blood samples when compared to commonly employed techniques. After upstream processing, isolated cells using OS were immunophenotyped to be CD14-, CD34-, CD45-, CD44+, CD105+, and CD106+, and displayed multipotent differentiation. Preliminary investigations to determine mechanisms responsible for osmolytic resistance revealed MSCs to have an ineffective volume of 59%, with the ability to double cell volume at infinite dilution. Disruption of filamentous actin polymerization by cytochalasin D sensitized MSCs to osmotic lysis, which suggests a cytoskeletal element involved in osmolytic resistance.</p>\",\"PeriodicalId\":23102,\"journal\":{\"name\":\"Tissue engineering\",\"volume\":\"13 10\",\"pages\":\"2465-73\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2007-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1089/ten.2007.0054\",\"citationCount\":\"28\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Tissue engineering\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1089/ten.2007.0054\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Tissue engineering","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/ten.2007.0054","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 28

摘要

未分化成体干细胞/祖细胞的分离仍然是一项具有挑战性的任务,主要是因为这些细胞在生物样品中数量稀少,并且缺乏独特的标记物。在此,我们报告了一种相对简单的分离人间充质干细胞(MSCs)的方法,这种方法基于它们对渗透裂解的异常抵抗,我们称之为“渗透选择”(OS)。MSCs可以显著地承受低渗条件(> 30分钟)的显著暴露,只有细胞增殖的可逆损伤,并且暴露后没有干细胞潜能的损失。MSCs与其他循环的非造血细胞的比较揭示了一个时间制度,通过该制度,这些细胞的纯化可以实现,而不会造成相当大的细胞损失。与常用的技术相比,OS显示脐带血样本中成纤维细胞集落形成单位的富集程度为50倍。经过上游处理后,使用OS分离的细胞免疫表型为CD14-、CD34-、CD45-、CD44+、CD105+和CD106+,并表现出多能分化。初步研究确定渗透抵抗机制的结果显示,MSCs的无效体积为59%,在无限稀释下能够使细胞体积翻倍。细胞松弛素D对丝状肌动蛋白聚合的破坏使间充质干细胞对渗透裂解敏感,这表明细胞骨架元件参与渗透抗性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Osmotic selection of human mesenchymal stem/progenitor cells from umbilical cord blood.

The isolation of undifferentiated adult stem/progenitor cells remains a challenging task primarily due to the rare quantity of these cells in biological samples and the lack of unique markers. Herein, we report a relatively straightforward method for isolation of human mesenchymal stem cells (MSCs) based on their unusual resistance to osmotic lysis, which we term "osmotic selection" (OS). MSCs can remarkably withstand significant exposure to hypotonic conditions (> 30 min) with only a reversible impairment in cell proliferation and with no loss of stem cell potential after exposure. Comparison of MSCs to other circulating nonhematopoietic cells revealed a time regime, by which purification of these cells would be attainable without considerable cell loss. OS showed a 50-fold enrichment of fibroblast colony-forming units from umbilical cord blood samples when compared to commonly employed techniques. After upstream processing, isolated cells using OS were immunophenotyped to be CD14-, CD34-, CD45-, CD44+, CD105+, and CD106+, and displayed multipotent differentiation. Preliminary investigations to determine mechanisms responsible for osmolytic resistance revealed MSCs to have an ineffective volume of 59%, with the ability to double cell volume at infinite dilution. Disruption of filamentous actin polymerization by cytochalasin D sensitized MSCs to osmotic lysis, which suggests a cytoskeletal element involved in osmolytic resistance.

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
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
自引率
0.00%
发文量
0
×
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:481959085
Book学术官方微信