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使用 Sepax® 技术验证一种自动化分离人脂肪组织来源细胞的程序。

Validation of an automated procedure to isolate human adipose tissue-derived cells by using the Sepax® technology.

机构信息

Department of Surgery, University Hospital Basel, Basel, Switzerland.

出版信息

Tissue Eng Part C Methods. 2012 Aug;18(8):575-82. doi: 10.1089/ten.TEC.2011.0617. Epub 2012 Apr 2.

Abstract

The stromal vascular fraction of adipose tissue has gained popularity as a source of autologous progenitor cells for tissue engineering and regenerative medicine applications. The aim of this study was to validate a newly developed, automated procedure to isolate adipose-derived mesenchymal stem/stromal cells (ASCs) from adult human lipoaspirates in a closed and clinical-grade device, based on the Sepax(®) technology. Using a total of 11 donors, this procedure was compared with the standard operator-based manual separation in terms of isolation yield, clonogenic fraction, phenotype, and differentiation potential of ASCs. As compared with the manual process, automation resulted in a 62% higher isolation yield, with 2.6±1.2×10(5) nucleated cells per mL of liposuction, and a 24% higher frequency of clonogenic progenitors. The variability in the isolation yield and clonogenicity across different preparations was reduced by 18% and 50%, respectively. The cytofluorimetric profile and in vitro differentiation capacity into mesenchymal lineages were comparable in the cells isolated using the two procedures. The new Sepax-based process thus allows an efficient isolation of ASCs with higher and more reproducible yields than the standard manual procedure, along with minimal operator intervention. These results are expected to facilitate the use of ASCs for clinical purposes, either within an intraoperative setting or in combination with further in vitro cell expansion/cultivation.

摘要

脂肪组织的基质血管部分作为组织工程和再生医学应用中自体祖细胞的来源已经越来越受到关注。本研究的目的是基于 Sepax(®)技术,验证一种新开发的、用于从成人脂肪抽吸物中分离脂肪来源间充质干细胞/基质细胞(ASCs)的自动化程序,该程序在封闭且临床级别的设备中进行。使用总共 11 个供体,将该程序与标准的基于操作者的手动分离方法在分离产量、集落形成部分、表型和 ASC 的分化潜能方面进行了比较。与手动过程相比,自动化操作可使分离产量提高 62%,每毫升脂肪抽吸物中含有 2.6±1.2×10(5)个有核细胞,集落形成祖细胞的频率提高 24%。两种方法的分离产量和克隆形成率的变异性分别降低了 18%和 50%。使用两种方法分离的细胞的细胞荧光分析图谱和体外向间充质谱系分化的能力相当。因此,新的基于 Sepax 的方法可有效分离 ASC,其产量更高、更具可重复性,且操作者干预最小。这些结果有望促进 ASC 在临床中的应用,无论是在术中环境中还是与进一步的体外细胞扩增/培养相结合。

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