Presentation: 2025 ND EPSCoR Annual conference 

October 21, 2025, NDSU Memorial Union, Fargo, North Dakota

Investigation of the Effect of Cell Surface Marker CD133 in Renal Progenitor Cells

The proximal tubule of the kidney is the main site of damage after exposure to toxins, hypoxia, or major injury. The renal tubular progenitor cell population is responsible for the recovery and regeneration of the proximal tubule after these events. These renal progenitor cells are sparsely scattered within the kidney and identified by their co-expression of the known stem cell markers CD133 and CD24. The hypothesis of this study is that CD133 plays a critical functional role in stemness and self-renewal of renal progenitor cells. Using CD24+/CD133+ renal progenitor (HRTPT) cells isolated from the immortalized RPTEC/TERT1 proximal tubular cell line, we established a CD133 knockdown with a lentiviral vector expressing short hairpin against CD133. We assessed gene and protein expression of CD133 and select stem cell markers using RT-qPCR and Simple Western procedures. We measured self-renewal capability by nephrosphere formation under ultralow-attachment conditions and multipotent differentiation capacity by cell growth under adipogenic, tubulogenic, osteogenic, and neurogenic conditions. Results showed that CD133 knockdown in HRTPT cells caused a significant decrease in the expression of stem cell markers (CD133, OCT4, SOX2, KLF4, CD44, and CMYC) and reduced self-renewal capacity as indicated by reduced nephrosphere formation. In addition, CD133 knockdown significantly decreased the differentiation capacity in tubulogenic and osteogenic media. Our results indicate that CD133 is critical for stem/progenitor characteristics in HRTPT cells and loss of CD133 results in decreased ability to perform self-renewal and multipotent differentiation capacity, indicating the importance of CD133 as a potential therapeutic target in acute kidney injury.