Tracking of Urine-Derived Stem Cells in Mice

Area of Science:

• Regenerative Medicine
• Cell Biology
• Biotechnology

Background:

• Urine-derived stem cells (USCs) are multipotent stem cells sourced noninvasively from human urine.
• USCs hold potential for autologous and allogeneic therapies, particularly for acute kidney injury (AKI) due to their renal origin.
• Previous studies have tracked USCs in mouse models of AKI.

Purpose of the Study:

• To outline a comprehensive protocol for isolating, culturing, and genetically modifying USCs for tracking.
• To detail the methodology for tracking USC migration and biodistribution in vivo using quantitative bioluminescence tomographic imaging (qBLT).
• To provide a precise method for studying cell localization in animal models.

Main Methods:

• Isolation and culture of human USCs.
• Transfection of USCs with luciferase piggyBac transposon plasmids for bioluminescence imaging.
• Administration of 1 × 10⁶ luciferase-labeled USCs (luc-USCs) via intraperitoneal injection in mouse models.
• Quantitative bioluminescence tomographic imaging (qBLT) for tracking cell migration and biodistribution.
• Generation of 3D bioluminescent images and data analysis using InVivoAX™ software.

Main Results:

• Established a detailed protocol for luc-USC preparation.
• Demonstrated the capability of qBLT to provide accurate spatial and temporal insights into USC migration.
• Successfully generated 3D bioluminescent images and analyzed cell localization data.

Conclusions:

• The presented protocol enables robust tracking of USCs in vivo.
• qBLT combined with luc-USCs offers a precise methodology for studying cell behavior in animal models.
• This approach facilitates further research into USC-based therapies for conditions like AKI.