Rapid Prototyping Laboratory, Department of Mechanical Engineering, School of Engineering, Stanford University, Stanford, California, USA.
Tissue Eng Part A. 2011 Feb;17(3-4):495-502. doi: 10.1089/ten.TEA.2010.0211. Epub 2010 Oct 25.
The recent technique of transducing key transcription factors into unipotent cells (fibroblasts) to generate pluripotent stem cells (induced pluripotent stem cells [iPSCs]) has significantly changed the stem cell field. These cells have great promise for many clinical applications, including that of regenerative medicine. Our findings show that iPSCs can be derived from human adipose-derived stromal cells (hASCs), a notable advancement in the clinical applicability of these cells. To investigate differences between two iPS cell lines (fibroblast-iPSC and hASC-iPSC), and also the gold standard human embryonic stem cell, we looked at cell stiffness as a possible indicator of cell differentiation-potential differences. We used atomic force microscopy as a tool to determine stem cell stiffness, and hence differences in material properties between cells. Human fibroblast and hASC stiffness was also ascertained for comparison. Interestingly, cells exhibited a noticeable difference in stiffness. From least to most stiff, the order of cell stiffness was as follows: hASC-iPSC, human embryonic stem cell, fibroblast-iPSC, fibroblasts, and, lastly, as the stiffest cell, hASC. In comparing hASC-iPSCs to their origin cell, the hASC, the reprogrammed cell is significantly less stiff, indicating that greater differentiation potentials may correlate with a lower cellular modulus. The stiffness differences are not dependent on cell culture density; hence, material differences between cells cannot be attributed solely to cell-cell constraints. The change in mechanical properties of the cells in response to reprogramming offers insight into how the cell interacts with its environment and might lend clues to how to efficiently reprogram cell populations as well as how to maintain their pluripotent state.
最近将关键转录因子转导为单能细胞(成纤维细胞)以生成多能干细胞(诱导多能干细胞[iPSCs])的技术极大地改变了干细胞领域。这些细胞在许多临床应用中具有很大的应用前景,包括再生医学。我们的研究结果表明,可以从人脂肪组织来源的基质细胞(hASCs)中获得 iPS 细胞,这是这些细胞在临床应用中的显著进步。为了研究两种 iPS 细胞系(成纤维细胞-iPSC 和 hASC-iPSC)之间的差异,以及黄金标准的人类胚胎干细胞,我们将细胞刚性作为细胞分化潜能差异的可能指标。我们使用原子力显微镜作为工具来确定干细胞的刚性,从而确定细胞之间在材料特性上的差异。还确定了人成纤维细胞和 hASC 的刚性以进行比较。有趣的是,细胞的刚性表现出明显的差异。细胞刚性从最低到最高的顺序如下:hASC-iPSC、人类胚胎干细胞、成纤维细胞-iPSC、成纤维细胞,最后是最硬的细胞 hASC。在将 hASC-iPSC 与其来源细胞 hASC 进行比较时,重编程细胞的刚性明显降低,这表明更大的分化潜能可能与较低的细胞模量相关。这些刚性差异不依赖于细胞培养密度;因此,细胞之间的材料差异不能仅仅归因于细胞间的限制。细胞在响应重编程时力学性能的变化提供了对细胞与环境相互作用方式的深入了解,并可能为有效重编程细胞群体以及如何维持其多能状态提供线索。