Abstract
A fundamental limitation in the derivation of hematopoietic stem and progenitor cells is the imprecise understanding of human developmental hematopoiesis. Herein we established a multilayer microfluidic Aorta-Gonad-Mesonephros (AGM)-on-a-chip to emulate developmental hematopoiesis from pluripotent stem cells. The device consists of two layers of microchannels separated by a semipermeable membrane, which allows the co-culture of human hemogenic endothelial (HE) cells and stromal cells in a physiological relevant spatial arrangement to replicate the structure of the AGM. HE cells derived from human induced pluripotent stem cells (hiPSCs) were cultured on a layer of mesenchymal stromal cells in the top channel while vascular endothelial cells were co-cultured on the bottom side of the membrane within the microfluidic device. We show that this AGM-on-a-chip efficiently derives endothelial-to-hematopoietic transition (EHT) from hiPSCs compared with regular suspension culture. The presence of mesenchymal stroma and endothelial cells renders functional HPCs in vitro. We propose that the AGM-on-a-chip could serve as a platform to dissect the cellular and molecular mechanisms of human developmental hematopoiesis.
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Acknowledgments
We would like to thank Dr. Misaki Ouchida for graphical assistance, Ms. Harumi Watanabe for administrative assistance, Ms. Yuka Ozaki and Kayo Yano for their technical assistance, and Dr. Peter Karagiannis for reading and editing the paper. This work was supported by the Core Center for iPS Cell Research of Research Center Network for Realization of Regenerative Medicine from the Japan Agency for Medical Research and Development (AMED) [M.K.S.], the Program for Intractable Diseases Research utilizing Disease-specific iPS cells (AMED: 17935423) [M.K.S.], and the Center for Innovation program of Japan Science and Technology Agency (JST) [R.O. and M.K.S.]. R.S. is a recipient of Early Career KAKENHI, iPS Academia Japan, and Sen-shin Medical Research Foundation (SMRF) fellowships. This work was also supported by AMED under Grant No. JP18gm5810008, JSPS KAKENHI Grant No. JP17H02082, and the Kyoto University Hakubi Project [Y.-s.T.].
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R.S., R.O. and Y-s.T. designed the study, conducted the experiments, interpreted the data and wrote the manuscript. C.M., A.L., T.M., E.S. and K.K. conducted the experiments. T.N. established the AGMS-3 cell line. A.N., M.K.S. and Y-s.T. supervised the study. T.N., A.N. and M.K.S. commented on and wrote the manuscript.
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Sugimura, R., Ohta, R., Mori, C. et al. Biomimetic aorta-gonad-mesonephros-on-a-chip to study human developmental hematopoiesis. Biomed Microdevices 22, 34 (2020). https://doi.org/10.1007/s10544-020-00488-2
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DOI: https://doi.org/10.1007/s10544-020-00488-2