Engineering Human Collecting System Reporter Pluripotent Stem Cell Lines

Key Personnel

Sanjay Jain (PI)
Washington University in St. Louis

  • Bendi Gong
    Washington University in St. Louis
  • Shondra Miller
    Washington University in St. Louis

Project Description


We propose to generate modified human pluripotent stem cell (hPSC) reporter lines with a genetically encoded activity sensor that tags key cell types important in development and function of the urinary collecting system. We will leverage an existing WTC11 hiPSC line that harbors GCaMP6 as an activity sensor thereby enhancing the functional utility of these lines by superimposing physiology onto development during or after a desired cell type or kidney organoid is formed. Multi-labeled reporter hiPSC cell lines of the collecting system in this proposal will enable RBK members to incorporate this essential component of the kidneys in efforts to optimize nephron formation, patterning and organization in scaffolds with the goal of making a kidney that maintains homeostasis and successfully expels urine.


  1. (Re)Building a Kidney.

    Oxburgh, L; Carroll, TJ; Cleaver, O; Gossett, DR; Hoshizaki, DK; Hubbell, JA; Humphreys, BD; Jain, S; Jensen, J; Kaplan, DL; Kesselman, C; Ketchum, CJ; Little, MH; McMahon, AP; Shankland, SJ; Spence, JR; Valerius, MT; Wertheim, JA; Wessely, O; Zheng, Y; Drummond, IA. J Am Soc Nephrol. 28(5):1370–1378. May 2017.

    (Re)Building a Kidney is a National Institute of Diabetes and Digestive and Kidney Diseases-led consortium to optimize approaches for the isolation, expansion, and differentiation of appropriate kidney cell types and the integration of these cells into complex structures that replicate human kidney function. The ultimate goals of the consortium are two-fold: to develop and implement strategies for in vitro engineering of replacement kidney tissue, and to devise strategies to stimulate regeneration of nephrons in situ to restore failing kidney function. Projects within the consortium will answer fundamental questions regarding human gene expression in the developing kidney, essential signaling crosstalk between distinct cell types of the developing kidney, how to derive the many cell types of the kidney through directed differentiation of human pluripotent stem cells, which bioengineering or scaffolding strategies have the most potential for kidney tissue formation, and basic parameters of the regenerative response to injury. As these projects progress, the consortium will incorporate systematic investigations in physiologic function of in vitro and in vivo differentiated kidney tissue, strategies for engraftment in experimental animals, and development of therapeutic approaches to activate innate reparative responses.