Description

Single-cell RNA-sequencing of pluripotent stem cell-derived kidney cells

Nancy Mah¹, Krithika Hariharan¹, Sujun Oh¹, Andreas Kurtz¹

¹Berlin-Brandenburg Center for Regenerative Therapies, Charite University Medicine, Augustenburger Platz 1, 13353 Berlin, Germany

Chronic kidney disease can lead to end stage renal disease, at which point there are only two life-saving options for the patient: kidney transplantation or life-long dialysis. Novel treatment options aim to regenerate the kidney by using the patient's own cells to derive new kidney cells. By using the patient's own urinary cells for the establishment of induced pluripotent stem cells (iPS), it is possible to maintain kidney-specific genotypes that may be causative for the renal disease phenotype. We are using a custom protocol that differentiates human iPS cells into specific renal cell types, which could be used to generate organoids for disease modelling. The transcriptome of the bulk differentiated cells at day 8 shows that many markers of the renal vesicle are expressed. In mammalian kidney development, the renal vesicle gives rise to podocytes, mesangial cells, and epithelial cells (proximal/distal tubule cells). Specific markers for these cell types are detected in the day 14, which points to a mixture of these cell types. To further examine the renal differentiation in detail, we have performed single-cell RNA-sequencing of iPS-derived renal cells at day 10. By classifying the potency of these cells to become specific renal types we hope to better understand the specification of renal cell types during this critical stage of nephron morphogenesis.