BIOS Lab On a Chip Group, MESA+ Institute for Nanotechnology, Max Planck - University of Twente Center for Complex Fluid Dynamics, University of Twente, Enschede, The Netherlands.
Applied Stem Cell Technologies, TechMed Centre, University of Twente, Enschede, The Netherlands.
Biomed Microdevices. 2021 May 31;23(2):30. doi: 10.1007/s10544-021-00556-1.
Human stem cell-derived cells and tissues hold considerable potential for applications in regenerative medicine, disease modeling and drug discovery. The generation, culture and differentiation of stem cells in low-volume, automated and parallelized microfluidic chips hold great promise to accelerate the research in this domain. Here, we show that we can differentiate human embryonic stem cells (hESCs) to early cardiac mesodermal cells in microfluidic chambers that have a volume of only 30 nanoliters, using discontinuous medium perfusion. 64 of these chambers were parallelized on a chip which contained integrated valves to spatiotemporally isolate the chambers and automate cell culture medium exchanges. To confirm cell pluripotency, we tracked hESC proliferation and immunostained the cells for pluripotency markers SOX2 and OCT3/4. During differentiation, we investigated the effect of different medium perfusion frequencies on cell reorganization and the expression of the early cardiac mesoderm reporter MESP1 by live-cell imaging. Our study demonstrates that microfluidic technology can be used to automatically culture, differentiate and study hESC in very low-volume culture chambers even without continuous medium perfusion. This result is an important step towards further automation and parallelization in stem cell technology.
人源干细胞衍生的细胞和组织在再生医学、疾病建模和药物发现方面具有巨大的应用潜力。在低体积、自动化和并行化的微流控芯片中生成、培养和分化干细胞，有望加速该领域的研究。在这里，我们展示了我们可以使用不连续的介质灌注，在仅 30 纳升体积的微流控腔室中将人胚胎干细胞（hESC）分化为早期心脏中胚层细胞。在一个包含集成阀的芯片上并行化了 64 个腔室，以时空隔离腔室并自动进行细胞培养基交换。为了确认细胞多能性，我们跟踪 hESC 的增殖，并通过免疫染色检测多能性标志物 SOX2 和 OCT3/4。在分化过程中，我们通过活细胞成像研究了不同介质灌注频率对细胞重排和早期心脏中胚层报告基因 MESP1 表达的影响。我们的研究表明，微流控技术可用于在非常低体积的培养腔室中自动培养、分化和研究 hESC，即使没有连续的介质灌注也是如此。这一结果是朝着进一步自动化和并行化干细胞技术迈出的重要一步。
