Human Adipose-Derived Stromal Cell Isolation Methods and Use in Osteogenic and Adipogenic In Vivo Applications
Elizabeth Brett, Ruth Tevlin, Adrian McArdle, Eun Young Seo, Charles K.F. Chan, Derrick C. Wan, Michael T. Longaker
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Abstract
Adipose tissue represents an abundant and easily accessible source of multipotent cells, which may serve as excellent building blocks for tissue engineering. This article presents a newly described protocol for isolating adipose-derived stromal cells (ASCs) from human lipoaspirate, compared to the standard protocol for harvesting ASCs established in 2001.
Human ASC isolation is performed using two methods, and resultant cells are compared through cell yield, cell viability, cell proliferation and regenerative potential. The osteogenic and adipogenic potential of ASCs isolated using both protocols are assessed in vitro and gene expression analysis is performed. The focus of this series of protocols is the regenerative potential of both cell populations in vivo . As such, the two in vivo animal models described are fat graft retention (soft tissue reconstruction) and calvarial defect healing (bone regeneration). The techniques described comprise fat grafting with cell assisted lipotransfer, and calvarial defect creation healed with cell-seeded scaffolds. © 2017 by John Wiley & Sons, Inc.
人脂肪来源的基质细胞分离方法及其在成骨和脂肪生成中的体内应用
脂肪组织是一种丰富且易于获取的多能细胞来源,可作为组织工程的优秀构建块。本文介绍了一种从人抽脂液中分离脂肪源性基质细胞(ASCs)的新方法,与2001年建立的脂肪源性基质细胞的标准方法进行了比较。采用两种方法分离人类ASC,并通过细胞产量、细胞活力、细胞增殖和再生潜能对分离得到的细胞进行比较。在体外评估两种方法分离的ASCs的成骨和成脂潜能,并进行基因表达分析。这一系列方案的重点是两种细胞群在体内的再生潜力。因此,描述的两种体内动物模型是脂肪移植物保留(软组织重建)和颅骨缺损愈合(骨再生)。所描述的技术包括细胞辅助脂肪移植的脂肪移植,以及细胞植入支架修复的颅骨缺损。©2017 by John Wiley &儿子,Inc。
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