# In Vivo Imaging of Immune Rejection of MIN6 Cells Transplanted in C3H Mice

## TL;DR

The study shows how immune rejection of transplanted MIN6 cells can be tracked noninvasively in mice using bioluminescence imaging.

## Contribution

The novel use of bioluminescence imaging to detect immune rejection of MIN6 cells in C3H mice is demonstrated.

## Key findings

- C3H mice showed immune rejection of MIN6 cells, as indicated by lack of hypoglycemia and bioluminescence signals.
- Presensitization with C57BL/6 spleen cells enhanced immune rejection in C3H mice.
- Bioluminescence imaging effectively detected rejection of subcutaneously transplanted MIN6 cells.

## Abstract

Recently, we successfully utilized noninvasive magnetic resonance and bioluminescence imaging to track MIN6 cells subcutaneously transplanted in immunocompromised nude mice for up to 64 days. In this study, we further used bioluminescence imaging to investigate the immune rejection of MIN6 cells in immunocompetent C3H mice. A total of 5 × 106 luciferase-transfected MIN6 cells were implanted into the subcutaneous space of each nude or C3H mouse. After transplantation, hypoglycemia and persistent bioluminescence signals were observed in eight of eight (100%) nude mice and five of nine (56%) C3H mice (p < 0.05). We then presensitized a group of C3H mice with C57BL/6 spleen cells just prior to transplantation (n = 14). Interestingly, none of them had hypoglycemia or persistent bioluminescence signals (p < 0.01 vs. C3H mice without presensitization). Histological examination of the grafts revealed a lack or minimal presence of insulin-positive cells in recipients without hypoglycemia and persistent bioluminescence signals. In contrast, recipients with hypoglycemia and persistent bioluminescence signals showed a significant presence of insulin-positive cells in their grafts. Our results indicate that rejection of MIN6 cells occurred in C3H mice and could be enhanced by presensitization with C57BL/6 spleen cells and that bioluminescence imaging is a useful noninvasive tool for detecting rejection of subcutaneously transplanted MIN6 cells.