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无酶技术可实现在床边大量分离脂肪来源干细胞。

An enzyme-free technique enables the isolation of a large number of adipose-derived stem cells at the bedside.

机构信息

Department of Medical Biology, Faculty of Medicine, Lokman Hekim University, Ankara, 06800, Turkey.

Plastic Reconstructive and Aesthetic Surgery, Lokman Hekim Hospital, Ankara, 06800, Turkey.

出版信息

Sci Rep. 2023 May 17;13(1):8005. doi: 10.1038/s41598-023-34915-0.

Abstract

Adipose tissue derived stromal cells (ADSCs) play a crucial role in research and applications of regenerative medicine because they can be rapidly isolated in high quantities. Nonetheless, their purity, pluripotency, differentiation capacity, and stem cell marker expression might vary greatly depending on technique and tools used for extraction and harvesting. There are two methods described in the literature for isolating regenerative cells from adipose tissue. The first technique is enzymatic digestion, which utilizes many enzymes to remove stem cells from the tissue they reside in. The second method involves separating the concentrated adipose tissue using non-enzymatic, mechanical separation methods. ADSCs are isolated from the stromal-vascular fraction (SVF) of processed lipoaspirate, which is the lipoaspirate's aqueous portion. The purpose of this work was to evaluate a unique device 'microlyzer' for generating SVF from adipose tissue using a mechanical technique that required minimal intervention. The Microlyzer was examined using tissue samples from ten different patients. The cells that were retrieved were characterized in terms of their cell survival, phenotype, proliferation capacity, and differentiation potential. The number of progenitor cells extracted only from the microlyzed tissue was in comparable amount to the number of progenitor cells acquired by the gold standard enzymatic approach. The cells that were collected from each group exhibit similar levels of viability as well as proliferation rates. In addition, the differentiation potentials of the cells derived from the microlyzed tissue were investigated, and it was discovered that cells isolated through microlyzer entered the differentiation pathways more quickly and displayed a greater level of marker gene expression than cells isolated by enzymatic methods. These findings suggest that microlyzer, particularly in regeneration investigations, will allow quick and high rate cell separation at the bedside.

摘要

脂肪组织来源的基质细胞(ADSCs)在再生医学的研究和应用中起着至关重要的作用,因为它们可以快速大量分离。然而,其纯度、多能性、分化能力和干细胞标志物表达可能因提取和收获所用的技术和工具而有很大差异。文献中描述了从脂肪组织中分离再生细胞的两种方法。第一种技术是酶消化,它利用多种酶从组织中去除干细胞。第二种方法涉及使用非酶、机械分离方法分离浓缩的脂肪组织。ADSCs 是从处理过的脂肪抽吸物的基质血管部分(SVF)中分离出来的,SVF 是脂肪抽吸物的水相部分。本工作的目的是评估一种独特的设备“微裂器”,该设备使用需要最小干预的机械技术从脂肪组织中产生 SVF。使用来自十个不同患者的组织样本检查了微裂器。从细胞存活、表型、增殖能力和分化潜能方面对回收的细胞进行了表征。仅从微裂组织中提取的祖细胞数量与通过金标准酶法获得的祖细胞数量相当。从每组收集的细胞表现出相似的存活率和增殖率。此外,还研究了源自微裂组织的细胞的分化潜力,发现通过微裂器分离的细胞比通过酶法分离的细胞更快地进入分化途径,并且表现出更高水平的标记基因表达。这些发现表明,微裂器,特别是在再生研究中,将允许在床边快速、高比例地分离细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a31a/10192379/6b9655ef5be3/41598_2023_34915_Fig1_HTML.jpg

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