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脂肪来源干细胞:两种临床应用分离方法的比较

Adipose-derived stem cells: Comparison between two methods of isolation for clinical applications.

作者信息

Raposio Edoardo, Simonacci Francesco, Perrotta Rosario E

机构信息

Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Parma, Italy.

Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Parma, Italy.

出版信息

Ann Med Surg (Lond). 2017 Jul 8;20:87-91. doi: 10.1016/j.amsu.2017.07.018. eCollection 2017 Aug.

Abstract

BACKGROUND

Adipose-derived stem cells are recognized as being an effective mesenchymal stem cell population with enormous potential in different fields of regenerative medicine and stem cell therapy. Although there is unanimous agreement on the harvesting procedure for adipose tissue, there are various protocols for adipose-derived stem cell isolation. The aim of this study was compare two methods of adipose-derived stem cells (ASCs) isolation, one based on a mechanical + enzymatic (ME) procedure and the other one exclusively mechanical (MC), in order to determine which one was superior to the other in accordance with current European and US legislation.

METHODS

We reported step by step the two different methods ASCs isolation by comparing them. The ME procedure included the use of a centrifuge, an incubator and collagenase digestion solution (Collagenase NB 6 GMP Grade 17458; Serva GmbH, Heidelberg, Germany). The MC procedure was performed by vibrating shaker and centrifuge, both placed in a laminar airflow bench.

RESULTS

With the ME procedure, a mean of 9.06 × 10 ASCs (range, 8.4 to 9.72 × 10; SD ± 6.6 × 10) was collected, corresponding to 25.9% of the total number of harvested cells. With the MC procedure, a mean of 5 × 10 ASCs (range: 4.0 to 6.0 × 10; SD, ±1 × 10) was collected, corresponding to 5% of the total number of harvested cells.

CONCLUSION

Based on data collected, from the same amount of lipoaspirate the ME procedure allowed to isolate a greater number of ASCs (25,9%) compared to the MC one (5%).

摘要

背景

脂肪来源干细胞被认为是一种有效的间充质干细胞群体,在再生医学和干细胞治疗的不同领域具有巨大潜力。尽管对于脂肪组织的采集程序已达成一致意见,但脂肪来源干细胞的分离方法却多种多样。本研究的目的是比较两种脂肪来源干细胞(ASC)的分离方法,一种基于机械+酶法(ME),另一种仅为机械法(MC),以便根据当前欧洲和美国的法规确定哪种方法更具优势。

方法

我们通过比较逐步报告了两种不同的ASC分离方法。ME方法包括使用离心机、培养箱和胶原酶消化液(胶原酶NB 6 GMP 17458级;德国海德堡的Serva GmbH公司)。MC方法通过置于层流通风柜中的振动摇床和离心机进行。

结果

采用ME方法,平均收集到9.06×10个ASC(范围为8.4至9.72×10;标准差±6.6×10),占收获细胞总数的25.9%。采用MC方法,平均收集到5×10个ASC(范围:4.0至6.0×10;标准差,±1×10),占收获细胞总数的5%。

结论

根据收集到的数据,从相同量的脂肪抽吸物中,与MC方法(5%)相比,ME方法能够分离出更多数量的ASC(25.9%)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c30f/5508488/f218f5feefae/gr1.jpg

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