Tracking Implanted DFAT Cells in Organs

DFAT cells from the ICR mice were labeled with PKH26 Red Fluorescent Cell Linker Kit for General Cell Membrane Labeling (Sigma Chemical, St. Louis, MO, USA) as described previously [13 (link)]. In total, 10⁵ labeled DFAT cells were infused through the posterior orbital venous plexus of the ICR mice. At 1 h, 24 h and 1 week after the injection, kidney, aorta, liver and lungs were removed and fixed in 3% formalin in phosphate-buffered saline (PBS) (Kanto Chemical, Tokyo, Japan) and embedded in paraffin. Sections were observed under a fluorescence microscope (IX73; Olympus, Tokyo, Japan), and images were obtained with a digital imaging system (Fig. 1).
Fig. 1

Distribution of implanted DFAT cells. In total, 10⁵ PKH26-labeled DFAT cells were infused through the posterior orbital venous plexus in ICR mice. One hour, 24 h, and one week after the infusion, kidney, aorta, liver and lungs were removed and fixed in 3% formalin in PBS and embedded in paraffin. Arrowheads indicate trapped DFAT cells. Bar = 50 μm. DFAT dedifferentiated fat