Abstract
In the future, stem cell-based technologies may be harnessed to replace conventional dialysis and transplantation in patients with diabetic nephropathy. Recently, there has been considerable effort to improve methods for the differentiation of human pluripotent stem cells (hPSCs) into kidney cells in culture. Here, we present a protocol for obtaining more advanced kidney structures than have currently been possible in vitro, including vascularized glomeruli and tubular elements. HPSCs are first differentiated in 2D culture to a kidney progenitor stage. These cells are then dissociated and injected subcutaneously into immunocompromised mice. Twelve weeks later, the cells have developed into mature kidney structures and are excised for further characterization. This method constitutes a significant improvement on protocols that involve either exclusively a 2D culture or placing the cells in 3D organoid culture at the air-liquid interface in vitro.
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Acknowledgments
We acknowledge grant support as follows: Research Councils UK/Medical Research Council (MRC) MR/K026739/1 UK Regenerative Medicine Platform Hub, Safety and Efficacy of Stem Cells; MRC MICA grant MR/M017344; Kidneys for Life pump priming grant KfL/1/15; Manchester Regenerative Medicine Network strategic funding initiative grant MARMN/1/17; and Kidney Research UK project grant JFS/RP/008/20160916.
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Bantounas, I., Silajdžić, E., Woolf, A.S., Kimber, S.J. (2020). Formation of Mature Nephrons by Implantation of Human Pluripotent Stem Cell-Derived Progenitors into Mice. In: Gnudi, L., Long, D. (eds) Diabetic Nephropathy. Methods in Molecular Biology, vol 2067. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9841-8_19
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DOI: https://doi.org/10.1007/978-1-4939-9841-8_19
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