Brett Elizabeth, Tevlin Ruth, McArdle Adrian, Seo Eun Young, Chan Charles K F, Wan Derrick C, Longaker Michael T
Technical University Munich, Department of Plastic and Hand Surgery, Munich, Germany.
Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Plastic and Reconstructive Surgery, Stanford University School of Medicine, Stanford, California, USA.
Curr Protoc Stem Cell Biol. 2017 Nov 15;43:2H.1.1-2H.1.15. doi: 10.1002/cpsc.41.
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.
脂肪组织是多能细胞的丰富且易于获取的来源，可作为组织工程的优质构建单元。本文介绍了一种新描述的从人抽脂物中分离脂肪来源基质细胞（ASC）的方案，并与2001年建立的收获ASC的标准方案进行了比较。使用两种方法进行人ASC分离，并通过细胞产量、细胞活力、细胞增殖和再生潜力对所得细胞进行比较。对使用两种方案分离的ASC的成骨和成脂潜力进行体外评估，并进行基因表达分析。这一系列方案的重点是两种细胞群体在体内的再生潜力。因此，所描述的两种体内动物模型是脂肪移植保留（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，以及用接种细胞的支架愈合颅骨缺损。© 2017约翰威立父子公司。
