脂肪干细胞分离、纯化和保存:研究进展与未来方向

Isolation, purification and preservation of adipose-derived stem cells:research progress and future development

背景:2001年,Zuk等从吸脂术抽出的脂肪中发现了脂肪干细胞,揭开了干细胞研究的新篇章。近年来的研究证实,干细胞广泛存在于体内各组织,但脂肪干细胞以其来源广泛、获取简单等优点,一直以来是整形修复科、组织工程和再生医学等相关学科的研究重点。目的:从脂肪组织的取材、脂肪干细胞的分离、纯化、传代培养和冻存等方面进行综述,讨论上述过程中影响脂肪干细胞产量、增殖和分化能力的主要因素,同时提出目前的问题和未来的研究方向。方法:于2015年9月10日在PubM ed中以(adipose stem cells[Title])OR(adipose-derived stem cells[Title])OR(adipose-derived mesenchymal stem cells[Title])作为检索式进行检索,在SinoM ed中以("脂肪干细胞"[中文标题:智能])or("脂肪间充质干细胞"[中文标题:智能])作为检索式进行检索。通过标题及摘要判断其主要内容,最终选取81篇代表性文献。文章还介绍了美国德克萨斯大学MD安德森肿瘤中心整形科和TRAMCEL实验室(Tissue Regeneration and Molecular Cell Engineering Lab)的相关经验。结果与结论:脂肪组织分布广泛,给脂肪干细胞提供了充足的来源。利用吸脂术获得脂肪组织,患者痛苦小,供区损伤低。胶原酶消化法操作简单,产量高,仍然是基础实验中脂肪干细胞的主要分离方法,但是临床应用中一般不分离脂肪干细胞,或者利用临床级的无异种胶原酶甚至无酶的分离方法。脂肪干细胞的产量、表型、增殖和分化能力等生物学特性会受到取材和分离过程中多种因素的影响,因此目前需要制定标准化的脂肪干细胞分离准则。

BACKGROUND： In 2001, Zuk et al found adipose-derived stem cells（ASCs） from the aspirate of liposuction for the first time, which launched a new era of stem cell research. In recent years, stem cells have been proved to widely exist in many tissues and organs. ASCs are always in the spotlight of plastic and reconstructive surgery, tissue engineering and regenerative medicine because of extensive sources and simple isolation. OBJECTIVE： To review the fat tissue harvesting and ASCs isolation, purification, expansion, and cryopreservation, to discuss the main factors which influence the yield, proliferation capacity and differentiation potential of ASCs, and to predict the future research interests based on current issues. METHODS： On September 10 th, 2015, relevant articles were searched in PubM ed using the following format：（adipose stem cells[Title]） OR（adipose-derived stem cells[Title]） OR（adipose-derived mesenchymal stem cells[Title]） and in Sino Med using the following format in Chinese：（＂adipose-derived stem cells＂ [Title]）or（＂adipose-derived mesenchymal stem cells＂[Title]）. Finally, 81 representative articles were included according to their titles and abstracts. In this review, we also introduced relevant experience about the aforementioned procedures from the Department of Plastic Surgery and Tissue Regeneration and Molecular Cell Engineering Lab of University of Texas, MD Anderson Cancer Center, USA. RESULTS AND CONCLUSION： The widely dispersed fat tissues potentially provide abundant stem cells for tissue engineering and regenerative medicine. Liposuction is a mini-invasive approach for harvesting fat tissues. Collagenase digestion is the major method for ASCs isolation due to its simplicity and high yield in basic research. However, clinical fat transplantation without ASCs isolation or non-collagenase isolation of stromal vascular fraction or ASCs is preferred. The phenotype, proliferation and differentiation capacity of ASCs may be affected by several factors during the fat tissue harvesting and ASCs isolation. Therefore, a standard protocol for ASCs isolation is needed.