Division of Nephrology, Department of Internal Medicine III, Goethe University, Frankfurt, Germany.
Cytotherapy. 2010;12(1):96-106. doi: 10.3109/14653240903377045.
The potential of cultured adipose-derived stem cells (ASC) in regenerative medicine and new cell therapeutic concepts has been shown recently by many investigations. However, while the method of isolation of ASC from liposuction aspirates depending on plastic adhesion is well established, a standard expansion medium optimally maintaining the undifferentiated state has not been described.
We cultured ASC in five commonly used culture media (two laboratory-made media and three commercially available media) and compared them with a standard medium. We analyzed the effects on cell morphology, proliferation, hepatocyte growth factor (HGF) expression, stem cell marker profile and differentiation potential. Proliferation was measured with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a fluorescent assay. Release of HGF was assessed by an immunoassay. Expression of characteristic stem cell-related transcription factors and markers was evaluated by quantitative polymerase chain reaction (qPCR) (Nanog, Sox-2, Rex-1, nestin and Oct-4) and flow cytometry (CD44, CD73, CD90, CD105 and CD166), and differentiation was shown by adipogenic medium.
The morphology and expansion of ASC were significantly affected by the media used, whereas none of the media influenced the ASC potential to differentiate into adipocytes. Furthermore, two of the media induced an increase in expression of transcription factors, an increased secretion of HGF and a decrease in CD105 expression.
Culture of ASC in one of these two media before using the cells in cell therapeutic approaches may have a benefit on their regenerative potential.
最近，许多研究表明，培养的脂肪来源干细胞（ASC）在再生医学和新的细胞治疗概念中具有潜力。然而，尽管从脂肪抽吸物中分离 ASC 的方法依赖于塑料黏附已得到很好的确立，但尚未描述最佳维持未分化状态的标准扩增培养基。
我们在五种常用的培养培养基（两种实验室自制培养基和三种市售培养基）中培养 ASC，并将其与标准培养基进行比较。我们分析了对细胞形态、增殖、肝细胞生长因子（HGF）表达、干细胞标志物谱和分化潜能的影响。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法和荧光测定法测量增殖。通过免疫测定法评估 HGF 的释放。通过定量聚合酶链反应（qPCR）（Nanog、Sox-2、Rex-1、巢蛋白和 Oct-4）和流式细胞术（CD44、CD73、CD90、CD105 和 CD166）评估特征性干细胞相关转录因子和标志物的表达，并通过脂肪生成培养基显示分化。
所用培养基显著影响 ASC 的形态和扩增，而没有一种培养基影响 ASC 分化为脂肪细胞的潜力。此外，两种培养基均诱导转录因子表达增加、HGF 分泌增加和 CD105 表达减少。
在将细胞用于细胞治疗方法之前，在这两种培养基中的一种中培养 ASC 可能有益于提高其再生潜力。
