Abstract: SA-PO430
Gene-Modified Urine-Derived Stem Cells Home to the Ischemic Kidney
Session Information
- Development and Regenerative Medicine
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 500 Development, Stem Cells, and Regenerative Medicine
Authors
- Woodard, Lauren Elizabeth, Vanderbilt University Medical Center/VA, Charleston, South Carolina, United States
- Welch, Richard C., Vanderbilt University Medical Center, Nashville, Tennessee, United States
- Williams, Felisha M., Vanderbilt University Medical Center, Nashville, Tennessee, United States
- Paragas, Neal, University of Washington, Seattle, Washington, United States
- Wilson, Matthew H., Vanderbilt University, Nashville, Tennessee, United States
Background
Acute kidney injury is a major cause of morbidity and mortality, with only half of those diagnosed surviving three months. The current therapies for acute kidney injury are supportive rather than regenerative. We sought to evaluate if human urine-derived stem cells were able to migrate to the kidney following ischemia/reperfusion injury in mice.
Methods
Urine is a practical and painless source of cells for gene and cell therapy applications. Urine-derived stem cells are adult human cells of renal origin that propagate in tissue culture in media containing growth factors on gelatin-coated plates. We have isolated, expanded, transfected, and tracked these cells following injection into live NSG immunocompromised mice order to assess their potential for regenerative gene and cell therapies.
Results
FACS characterization revealed that they expressed the characteristic marker panel (CD44, CD73, CD90, & CD146 + / CD31, CD34, & CD45 -). They differentiated into osteogenic and adipogenic lineages. Transfection was optimized to achieve 61% transfection. Five different piggyBac luciferase transposons were compared with the CMV and EF1-alpha giving the highest luciferase signals. When luciferase-modified urine-derived stem cells were injected directly into the renal pelvis of immunocompromised NSG mice they migrated to the surgical wound scar. To test the homing ability of the cells in the setting of ischemia reperfusion injury, we optimized the time of ischemia together with unilateral nephrectomy in immunocompromised NSG mice to be 22 minutes with 83% of mice having elevated creatinine. Urine-derived stem cells transfected with piggyBac transposons expressing luciferase were injected into the peritoneum of mice on Day 3 post-ischemia reperfusion injury. The cells were tracked via the InVivoPLOT imaging gantry for quantitative tomographic optical live animal imaging. We found luciferase signal localized within 2 hours post-injection to the injured kidney with lower levels in the spleen.
Conclusion
Urine-derived stem cells represent an easily isolated, clinically relevant cell type that can be manipulated with non-viral genetic tools. We have found that the cells quickly migrate from their injection site to injured tissues. Next we will assess functional correction following injection of urine-derived stem cells into mouse models of acute kidney injury.
Funding
- NIDDK Support