Tracking of Transplanted Stem Cells in Mouse Models Using Practical Fluorescence-Based Protocols

Methods Mol Biol. 2026 May 8. doi: 10.1007/7651_2026_704. Online ahead of print.

Abstract

Precise tracking of transplanted cells within mouse tissues is essential for evaluating engraftment, migration, and long-term persistence in transplantation and regenerative medicine research. Here, we present a practical and versatile cell-tracking workflow focusing on PKH26-based direct fluorescent labeling and GFP-mediated indirect genetic labeling. PKH26 provides rapid and robust membrane labeling that is useful for short-term tracing, while GFP enables stable and heritable fluorescence suitable for long-term monitoring of transplanted cells. This chapter provides optimized protocols, troubleshooting strategies, and critical technical considerations for implementing these complementary methods. Together, these approaches offer a flexible and reliable framework for evaluating the engraftment, survival, distribution, and behavior of transplanted cells in mouse models.

Keywords: Allogeneic transplantation; Direct fluorescent labeling; Indirect genetic labeling; Stem cell tracking; Xenotransplantation.