Figure 5.

Transplantation of lentivirus‐transduced HAM cells into mice. (a) Flow cytometry confirmed high transduction rate of EGFP lentivirus in HAM cells. More than 98% of cells show EGFP expression. (b) Fluorescence microscopic image of HAM cells expressing EGFP. (c) Transplantation of EGFP expressing HAM cells into immunocompetent CD‐1 mice by intrauterine infusion. Small volume of EGFP‐HAM cells at 1 × 10⁵ cells/0.1 mL were infused into the uterine lumen. Uteri were taken after 10 days and were frozen for cryosectioning. Sections were stained with Hoechst dye after fixation. Image on the far left shows nuclear staining (blue) and green fluorescence in the middle indicates EGFP fluorescence. These images are overlaid and shown on the right. (d) Intravenous infusion of EGFP‐HAM cells was performed via tail veins of anaesthetized nude mice. About 200 µL of cells were given at 5 × 10⁴ cells/0.1 mL. Tissues were collected for up to 4 weeks at 1‐week interval and frozen for cryosectioning. This representative figure is obtained from lung sections from a nude mouse at 4 weeks after infusion. Presence of EGFP‐HAM cells was again confirmed by immunofluorescence imaging using human specific anti‐CD59 or anti‐CD105 antibodies. Control is a lung section from a non‐injected mouse. Laser confocal microscopic images are shown at 60×. Insets are shown enlarged in the right panels [EGFP alone, EGFP (green) + CD marker (red) overlay].