机械分离的基质血管成分提供了一种有效的无胶原酶替代技术：一项比较研究。
Mechanically Isolated Stromal Vascular Fraction Provides a Valid and Useful Collagenase-Free Alternative Technique: A Comparative Study.
作者信息
Chaput Benoit, Bertheuil Nicolas, Escubes Marina, Grolleau Jean-Louis, Garrido Ignacio, Laloze Jérôme, Espagnolle Nicolas, Casteilla Louis, Sensebé Luc, Varin Audrey
机构信息
Toulouse, Rennes, and Limoges, France.
From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
出版信息
Plast Reconstr Surg. 2016 Oct;138(4):807-819. doi: 10.1097/PRS.0000000000002494.
BACKGROUND
The use of stromal vascular fraction and adipose-derived stromal cells in tissue regeneration is now being increasingly investigated, and studies have demonstrated that adipose-derived stromal cells present differentiation and immunomodulatory capacities. The development of a rapid, inexpensive, and enzyme-free technique to isolate adipose-derived stromal cell-enriched stromal vascular fraction is a major goal for stem cell therapy. Therefore, the authors compared innovative mechanical procedures to the gold standard technique, collagenase digestion.
METHODS
Stromal vascular fraction was prepared from 21 liposuctions using either enzymatic digestion or two different mechanical methods: high vortexing/centrifugation and dissociation by intersyringe processing. The effects of tissue processing on cell count, viability, proliferation, clonogenic enrichment, and the phenotypes of the different native cell were determined. Adipose-derived stromal cell phenotypes from the different protocols, and their differentiation and immunosuppressive potential, were compared.
RESULTS
Enzymatic digestion isolated more viable cells than dissociation by intersyringe processing and vortexing/centrifugation. The expansion rate and clonogenic enrichment were higher for stromal vascular fraction isolated with collagenase. The proportion of adipose-derived stromal cells was higher in stromal vascular fraction extracted with dissociation than with enzymatic digestion and vortexing/centrifugation (p < 0.01). Interestingly, all cultured adipose-derived stromal cells displayed similar differentiation and immunosuppressive capacities.
CONCLUSIONS
Enzymatic digestion extracts more adipose-derived stromal cells, but intersyringe dissociation enables the rapid extraction of adipose-derived stromal cell-enriched stromal vascular fraction. Moreover, mechanical methods enable adipose-derived stromal cell isolation with stemness and immunosuppressive properties, similar to enzymatic digestion. Such mechanical procedures could allow easier and more rapid isolation of adipose-derived stromal cell-enriched stromal vascular fraction for practitioners.
CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.
背景
目前，基质血管成分和脂肪来源的间充质干细胞在组织再生中的应用正受到越来越多的研究，并且研究表明脂肪来源的间充质干细胞具有分化和免疫调节能力。开发一种快速、廉价且无酶的技术来分离富含脂肪来源间充质干细胞的基质血管成分是干细胞治疗的一个主要目标。因此，作者将创新的机械方法与金标准技术胶原酶消化法进行了比较。
方法
使用酶消化法或两种不同的机械方法（高涡旋/离心法和注射器间处理解离法）从21例抽脂术中制备基质血管成分。确定了组织处理对细胞计数、活力、增殖、克隆形成富集以及不同天然细胞表型的影响。比较了不同方案中脂肪来源间充质干细胞的表型及其分化和免疫抑制潜力。
结果
酶消化法分离出的活细胞比注射器间处理解离法和涡旋/离心法更多。用胶原酶分离的基质血管成分的扩增率和克隆形成富集率更高。与酶消化法和涡旋/离心法相比，用注射器间处理解离法提取的基质血管成分中脂肪来源间充质干细胞的比例更高（p < 0.01）。有趣的是，所有培养的脂肪来源间充质干细胞都表现出相似的分化和免疫抑制能力。
结论
酶消化法能提取更多的脂肪来源间充质干细胞，但注射器间解离法能快速提取富含脂肪来源间充质干细胞的基质血管成分。此外，机械方法能够分离出具有干性和免疫抑制特性的脂肪来源间充质干细胞，类似于酶消化法。这种机械方法可为从业者更轻松、快速地分离富含脂肪来源间充质干细胞的基质血管成分。
临床问题/证据水平：治疗性，V级
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