Abstract
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 MESP1mCherry 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.
Highlights
Stem cell technology advances the generation and controlled differentiation of stem cells, including human adult stem cells, human embryonic stem cells and human induced pluripotent stem cells
We focus on validating our chip for culturing and differentiating human embryonic stem cells (hESCs) to cardiac mesodermal cells as the first step towards addressing this complex problem
In this article we showed that we can differentiate hESCs to cardiac mesoderm cells in 30 nL microchambers using different discontinuous medium perfusion intervals
Summary
Stem cell technology advances the generation and controlled differentiation of stem cells, including human adult stem cells, human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Stem cells have enabled new strategies for use in fields like regenerative medicine, disease modeling and drug discovery. Stem cell technology enables the Andries D. van der Meer and Mathieu Odijk have contributed to this work. Directing stem cell fate of human pluripotent stem cells (hPSCs) is essentially achieved by emulating embryogenesis and requires creating a highly dynamic and spatially heterogenous microenvironment. Reliably and reproducibly generating cell lineages with a high degree of cell commitment presents an enormous challenge. Significant progress has been made to overcome this challenge by
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