Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Poland.
Analyst. 2020 Apr 21;145(8):3017-3028. doi: 10.1039/c9an02366f. Epub 2020 Mar 5.
Stem cells (SCs) are more and more often applied in tissue engineering and cell therapies, e.g. in regenerative medicine. Standard methods of SC differentiation are time consuming and ineffective. Therefore, new bioanalytical methods (i.e. Lab-on-a-Chip systems) are develop to improve such type of studies. Although, microtechnology is a rapidly growing research area, there are so far not too many works which present SC differentiation into cardiomyocytes in the microsystems. Therefore, we present new microbioanalytical method of SC differentiation towards cardiac cells using a newly developed digitally controlled microdispenser integrated with a Heart-on-a-chip system. Seven-day culture of human mesenchymal stem cells (hMSCs) and their differentiation using biochemical factors such as 5-AZA (2 μM, 24 h) and VEGF (20 ng ml, 72 h) were investigated in the microsystem which was automatically operated using smartphone software. hMSC differentiation into the cardiac cells was confirmed using immunostaining of cardiac markers (α-actinin and troponin T). The usage of the microsystem allowed shortening the time of hMSC differentiation in comparison to macroscale method. We showed that the microsystem, in which the in vivo microenvironment is mimicked and dynamic conditions are provided by a microdispenser, favorably affect hMSC differentiation towards cardiac cells. Based on the presented research we can conclude that the developed digitally controlled microsystem could be successfully utilized as a new microbioanalytical method for stem cells differentiation and analysis of their function under dynamic conditions. In the future, this could be a helpful tool for scientists working on regenerative medicine.
干细胞(SCs)越来越多地应用于组织工程和细胞治疗,例如再生医学。SCs 的标准分化方法既耗时又低效。因此,新的生物分析方法(即芯片实验室系统)被开发出来以改进此类研究。尽管微技术是一个快速发展的研究领域,但到目前为止,关于微系统中 SC 向心肌细胞分化的研究还不多。因此,我们提出了一种新的微生物分析方法,使用新开发的数字控制微分配器与心脏芯片系统集成,将 SC 分化为心肌细胞。在微系统中研究了人骨髓间充质干细胞(hMSCs)的 7 天培养及其使用生化因子(如 5-AZA(2 μM,24 h)和 VEGF(20 ng ml,72 h)的分化,该微系统使用智能手机软件自动操作。通过心肌标志物(α-肌动蛋白和肌钙蛋白 T)的免疫染色证实 hMSC 向心肌细胞分化。与宏观方法相比,微系统的使用缩短了 hMSC 分化的时间。我们表明,微系统模拟了体内微环境,并通过微分配器提供动态条件,有利于 hMSC 向心肌细胞分化。基于所提出的研究,我们可以得出结论,开发的数字控制微系统可以成功用作新的微生物分析方法,用于干细胞分化和在动态条件下分析其功能。在未来,这可能是从事再生医学的科学家的有用工具。