TY - JOUR
T1 - Highly parallelized human embryonic stem cell differentiation to cardiac mesoderm in nanoliter chambers on a microfluidic chip
AU - Vollertsen, Anke R.
AU - Den, Simone A. ten
AU - Schwach, Verena
AU - van den Berg, Albert
AU - Passier, Robert
AU - van der Meer, Andries D.
AU - Odijk, Mathieu
N1 - Funding Information:
This work was supported by the VESCEL ERC Advanced Grant to A. van den Berg (Grant no. 669768) and by the Netherlands Organ-on-Chip Initiative (NOCI), an NWO Gravitation Project funded by the Ministry of Education, Culture and Science of the Government of the Netherlands, under Grant 024.003.001. The authors also thank Arnoud Onnink for valuable discussions on data analysis.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/6
Y1 - 2021/6
N2 - 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.
AB - 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.
KW - UT-Hybrid-D
KW - Human pluripotent stem cells
KW - Cardiac mesoderm
KW - Parallelization
KW - Discontinuous perfusion
KW - Microfluidic large-scale integration
UR - http://www.scopus.com/inward/record.url?scp=85107387347&partnerID=8YFLogxK
U2 - 10.1007/s10544-021-00556-1
DO - 10.1007/s10544-021-00556-1
M3 - Article
SN - 1387-2176
VL - 23
JO - Biomedical microdevices
JF - Biomedical microdevices
IS - 2
M1 - 30
ER -