Maioli Margherita, Rinaldi Salvatore, Santaniello Sara, Castagna Alessandro, Pigliaru Gianfranco, Delitala Alessandro, Bianchi Francesca, Tremolada Carlo, Fontani Vania, Ventura Carlo
Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
Cell Transplant. 2014;23(12):1489-500. doi: 10.3727/096368913X672037. Epub 2013 Aug 30.
Human adipose-derived stem cells (hASCs) have been recently proposed as a suitable tool for regenerative therapies for their simple isolation procedure and high proliferative capability in culture. Although hASCs can be committed into different lineages in vitro, the differentiation is a low-yield and often incomplete process. We have recently developed a novel nonenzymatic method and device, named Lipogems, to obtain a fat tissue derivative highly enriched in pericytes/mesenchymal stem cells by mild mechanical forces from human lipoaspirates. When compared to enzymatically dissociated cells, Lipogems-derived hASCs exhibited enhanced transcription of vasculogenic genes in response to provasculogenic molecules, suggesting that these cells may be amenable for further optimization of their multipotency. Here we exposed Lipogems-derived hASCs to a radioelectric asymmetric conveyer (REAC), an innovative device asymmetrically conveying radioelectric fields, affording both enhanced differentiating profiles in mouse embryonic stem cells and efficient direct multilineage reprogramming in human skin fibroblasts. We show that specific REAC exposure remarkably enhanced the transcription of prodynorphin, GATA-4, Nkx-2.5, VEGF, HGF, vWF, neurogenin-1, and myoD, indicating the commitment toward cardiac, vascular, neuronal, and skeletal muscle lineages, as inferred by the overexpression of a program of targeted marker proteins. REAC exposure also finely tuned the expression of stemness-related genes, including NANOG, SOX-2, and OCT-4. Noteworthy, the REAC-induced responses were fashioned at a significantly higher extent in Lipogems-derived than in enzymatically dissociated hASCs. Therefore, REAC-mediated interplay between radioelectric asymmetrically conveyed fields and Lipogems-derived hASCs appears to involve the generation of an ideal "milieu" to optimize multipotency expression from human adult stem cells in view of potential improvement of future cell therapy efforts.
人脂肪来源干细胞（hASCs）因其分离过程简单且在培养中具有高增殖能力，最近被认为是再生疗法的合适工具。尽管hASCs在体外可分化为不同谱系，但这种分化是一个低产且往往不完全的过程。我们最近开发了一种名为Lipogems的新型非酶法和装置，通过对人抽脂物施加温和机械力，获得富含周细胞/间充质干细胞的脂肪组织衍生物。与酶解细胞相比，Lipogems来源的hASCs在对促血管生成分子的反应中表现出血管生成基因转录增强，表明这些细胞可能适合进一步优化其多能性。在此，我们将Lipogems来源的hASCs暴露于一种射频不对称输送器（REAC），这是一种不对称输送射频电场的创新装置，它既能增强小鼠胚胎干细胞的分化特征，又能在人皮肤成纤维细胞中实现高效的直接多谱系重编程。我们发现，特定的REAC暴露显著增强了强啡肽原、GATA-4、Nkx-2.5、VEGF、HGF、vWF、神经生成素-1和肌分化抗原（myoD）的转录，这表明通过靶向标记蛋白程序的过表达推断其向心脏、血管、神经元和骨骼肌谱系的分化。REAC暴露还精细调节了干性相关基因的表达，包括NANOG、SOX-2和OCT-4。值得注意的是，REAC诱导的反应在Lipogems来源的hASCs中比在酶解的hASCs中显著更高程度地形成。因此，鉴于未来细胞治疗努力的潜在改善，REAC介导射频不对称输送电场与Lipogems来源的hASCs之间的相互作用似乎涉及产生一个理想的“环境”来优化成人干细胞的多能性表达。
