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{"title":"Isolation of Ready-to-Use Adipose-Derived Stem Cell (ASC) Pellet for Clinical Applications and a Comparative Overview of Alternate Methods for ASC Isolation","authors":"Edoardo Raposio, Nicolò Bertozzi","doi":"10.1002/cpsc.29","DOIUrl":null,"url":null,"abstract":"<p>Current literature does not offer a standardized method to isolate adipose-derived stem cells (ASCs) for clinical applications and hence clinical studies using ASCs often show inconsistent results. Most of these studies borrow laboratory or benchside-derived protocols, which are complex, time consuming, and involve the use of chemical, animal-derived reagents. In this unit we describe a relatively simple and faster isolation protocol that allows collection of a ready-to-use ASC pellet for clinical application. All steps are performed in a closed circuit in order to guarantee maximum process sterility. Once the adipose tissue is harvested by means of a standard liposuction procedure, it undergoes a first centrifugation in order to remove the oil and serous fractions. Then ASCs are released by enzymatic digestion from the surrounding connective tissue scaffold. Finally a double series of washing and centrifugation allows one to obtain the ASC pellet alone. We usually graft this ASC pellet onto the skin edge and to the bottom of chronic skin ulcers as ASCs proved to be effective in promoting wound healing processes. Moreover, an increasing number of clinical studies are currently ongoing to test their potential in every medical field, from orthopedics to cardiology, oncology, autoimmune diseases, and tissue engineering. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":"41 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.29","citationCount":"41","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Stem Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpsc.29","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 41
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Abstract
Current literature does not offer a standardized method to isolate adipose-derived stem cells (ASCs) for clinical applications and hence clinical studies using ASCs often show inconsistent results. Most of these studies borrow laboratory or benchside-derived protocols, which are complex, time consuming, and involve the use of chemical, animal-derived reagents. In this unit we describe a relatively simple and faster isolation protocol that allows collection of a ready-to-use ASC pellet for clinical application. All steps are performed in a closed circuit in order to guarantee maximum process sterility. Once the adipose tissue is harvested by means of a standard liposuction procedure, it undergoes a first centrifugation in order to remove the oil and serous fractions. Then ASCs are released by enzymatic digestion from the surrounding connective tissue scaffold. Finally a double series of washing and centrifugation allows one to obtain the ASC pellet alone. We usually graft this ASC pellet onto the skin edge and to the bottom of chronic skin ulcers as ASCs proved to be effective in promoting wound healing processes. Moreover, an increasing number of clinical studies are currently ongoing to test their potential in every medical field, from orthopedics to cardiology, oncology, autoimmune diseases, and tissue engineering. © 2017 by John Wiley & Sons, Inc.
用于临床应用的即食脂肪源性干细胞(ASC)颗粒的分离和ASC分离替代方法的比较综述
目前的文献没有提供一种标准化的方法来分离用于临床应用的脂肪源性干细胞(ASCs),因此使用ASCs进行的临床研究往往显示出不一致的结果。这些研究大多采用实验室或台式衍生方案,这些方案复杂、耗时,并涉及使用化学动物衍生试剂。在本单元中,我们描述了一个相对简单和快速的分离方案,允许收集准备使用的ASC颗粒用于临床应用。所有步骤都在封闭回路中进行,以保证最大限度的工艺无菌性。一旦脂肪组织通过标准吸脂程序被收集,它将经历第一次离心,以去除油和浆液部分。然后,ASCs通过酶消化从周围结缔组织支架中释放出来。最后,双系列洗涤和离心可以单独获得ASC颗粒。我们通常将ASC颗粒移植到慢性皮肤溃疡的皮肤边缘和底部,因为ASC被证明能有效促进伤口愈合过程。此外,目前正在进行越来越多的临床研究,以测试它们在各个医学领域的潜力,从骨科到心脏病学、肿瘤学、自身免疫性疾病和组织工程。©2017 by John Wiley &儿子,Inc。
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