Schneider Sandra, Unger Marina, van Griensven Martijn, Balmayor Elizabeth R
Experimental Trauma Surgery, Department of Trauma Surgery, Klinikum rechts der Isar, Technical University of Munich, Ismaninger Strasse 22, 81675, Munich, Germany.
Eur J Med Res. 2017 May 19;22(1):17. doi: 10.1186/s40001-017-0258-9.
The use of mesenchymal stem cells (MSCs) in research and in regenerative medicine has progressed. Bone marrow as a source has drawbacks because of subsequent morbidities. An easily accessible and valuable source is adipose tissue. This type of tissue contains a high number of MSCs, and obtaining higher quantities of tissue is more feasible. Fat tissue can be harvested using different methods such as liposuction and resection. First, a detailed isolation protocol with complete characterization is described. This also includes highlighting problems and pitfalls. Furthermore, some comparisons of these different harvesting methods exist. However, the later characterization of the cells is conducted poorly in most cases.
We performed an in-depth characterization over five passages including an investigation of the effect of freezing and thawing. Characterization was performed using flow cytometry with CD markers, metabolic activity with Alamar Blue, growth potential in between passages, and cytoskeleton staining.
Our results show that the cells isolated with distinct isolation methods (solid versus liposuction "liquid") have the same MSC potential. However, the percentage of cells positive for the markers CD73, CD90, and CD105 is initially quite low. The cells isolated from the liquid fat tissue grow faster at higher passages, and significantly more cells display MSC markers.
In summary, we show a simple and efficient method to isolate adipose-derived mesenchymal stem cells from different preparations. Liposuctions and resection can be used, whereas liposuction has more growth potential at higher passages.
间充质干细胞(MSCs)在研究和再生医学中的应用不断发展。骨髓作为来源存在缺点,因为会引发后续疾病。一个易于获取且有价值的来源是脂肪组织。这种组织含有大量间充质干细胞,获取更多数量的组织更可行。脂肪组织可以通过抽脂和切除等不同方法获取。首先,描述了一种具有完整特性的详细分离方案。这还包括突出问题和陷阱。此外,对这些不同获取方法存在一些比较。然而,在大多数情况下,细胞的后续特性鉴定做得很差。
我们对传代五次的细胞进行了深入特性鉴定,包括对冻融效果的研究。使用带有CD标记的流式细胞术、用阿拉玛蓝检测代谢活性、传代期间的生长潜力以及细胞骨架染色进行特性鉴定。
我们的结果表明,用不同分离方法(固体与抽脂“液体”)分离的细胞具有相同的间充质干细胞潜能。然而,标记物CD73、CD90和CD105阳性的细胞百分比最初相当低。从液体脂肪组织分离的细胞在较高传代时生长更快,并且有更多细胞显示出间充质干细胞标记物。
总之,我们展示了一种从不同制剂中分离脂肪来源间充质干细胞的简单有效方法。可以使用抽脂和切除方法,而抽脂在较高传代时有更大的生长潜力。