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Abstract: FR-PO774

Percutaneous Intrarenal Transplantation of Differentiated Induced Pluripotent Stem Cells into Newborn Mice

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Lau, Ricky Wai Kiu, Monash University, Clayton, Victoria, Australia
  • Al-Rubaie, Ali H., Monash University, Clayton, Victoria, Australia
  • Tripathi, Pratibha, Monash University, Melbourne, New South Wales, Australia
  • Ricardo, Sharon D., Monash University, Clayton, Victoria, Australia
Background

The reprogramming of adult cells to generate iPSCs may provide an unprecedented opportunity to elucidate disease mechanisms in vitro; for disease-modifying bioassays, and to advance cell-replacement therapy. The in vivo engraftment and survival of patient-derived iPSCs following allogenic transplantation into host kidneys remains a challenge. Here we investigate the survival and engraftment of human dermal derived iPSCs using a newborn mouse model, which represents a receptive immuno-privileged host environment.

Methods

iPSCs were generated from skin biopsies of patients using Sendai virus reprogramming. Differentiation of iPSCs to podocytes (iPSC-POD) was performed in Nphs1 (Nephrin)-green fluorescent protein (GFP) iPSCs by the addition of activin A, bone morphogenetic protein 7 (BMP7) and retinoic acid over 15 days of culture. To assess in vivo integration, undifferentiated iPSCs and iPSC-POD differentiated for 10 or 15 days were labeled with either carboxyfluorescein succinimidyl ester (CFSE) or long term cell tracking Qtracker® 705 probe. Thereafter, 1 x 105 of differentiated iPSC-POD were resuspended and injected in 10 ul of phosphate buffered saline solution directly into the kidneys of mouse pups at postnatal day one (P1). A timecourse analysis of cell integration was assessed in differentiated iPSC-POD, compared to undifferentiated cells, using confocal fluorescence microscopy and the co-expression of glomerular and podocyte-specific markers.

Results

D10 differentiated iPSC-PODs, that were positive for podocyte markers including podocin, were detected following direct kidney injection in newborn mice after P3. Undifferentiated iPSC-PODs were not detected at the same timepoint. The transplanted cells were viable and located in the developing kidney cortex including the tubulointerstitium and localised adjacent to developing glomeruli in the outer nephrogenic zone where they were found to co-localise with glomerular-specific markers including podocin, synaptopodin and Wilms tumour 1 (WT1).

Conclusion

This study provides proof-of-principle that transplanted differentiated iPSC–POD can survive and integrate into recipient newborn mouse kidneys, and represents an ideal in vivo environment due to the immature and immunoprivileged nature of the developing postnatal kidneys.

Funding

  • Private Foundation Support