Related Experiment Video
Updated: May 28, 2026
Differentiating Functional Roles of Gene Expression from Immune and Non-immune Cells in Mouse Colitis by Bone Marrow Transplantation
Published on: October 1, 2012
Bone marrow cells in murine colitis: multi-signal analysis confirms pericryptal myofibroblast engraftment without
Chung-Yin Lee1, Rosemary Jeffery, Gillian Hutchinson
1Histopathology Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
Background:
The contribution of bone marrow-derived cells to epithelial tissues in the inflamed gut remains controversial. Recent reports have suggested that cell fusion between bone marrow-derived cells and the intestinal epithelium takes place in inflammatory conditions.
Methods:
In attempts to confirm this, we have undertaken gender mis-matched bone marrow (BM) transplants from male Swiss Webster (SWR) mice to B and T cell-deficient female Rag2 KO mice which, 4 weeks later, were given 5% dextran sodium sulphate in drinking water to induce acute colitis. A further BM-treated group of animals with a graft versus host-like condition was also studied. We developed a new method to combine up to three brightfield or fluorescent lectin- or immuno-histochemical signals with fluorescent in situ hybridisation for the Y and X chromosomes to enable us unequivocally to identify BM-derived male cells which presented as different cell types in the gastrointestinal tract.
Principal Findings:
In rolled preparations of whole intestines we scanned around 1.5 million crypts at many tissue levels. In no instance did we see a Y chromosome-positive cell in the epithelial compartment, which was not also CD45-positive. We saw no evidence of cell fusion, based on combined X and Y chromosome analysis. Levels of CD45-positive stromal and lymphoid cells and pericryptal myfibroblasts (positive for α-smooth muscle actin) increased with time up to a plateau, which resembled the level seen in untreated control grafted animals. We saw very few Y chromosome-positive endothelial cells in intestinal stromal vessels.
Conclusions:
We conclude that whole BM transplantation does not result in intestinal epithelial engraftment in this model. Our new methods can usefully assist in multi-signal analyses of cell phenotypes following BM transplant and in models of chimaerism and regenerative medicine.
Insights
Bone marrow transplantation does not lead to intestinal epithelial engraftment in inflamed guts. New methods were developed to track bone marrow-derived cells, confirming no cell fusion occurred.
Area of Science:
- Gastroenterology
- Immunology
- Regenerative Medicine
Background:
- The role of bone marrow-derived cells in the inflamed gut epithelium is debated.
- Previous studies suggested cell fusion between bone marrow and intestinal cells during inflammation.
Purpose of the Study:
- To investigate bone marrow-derived cell contribution to the intestinal epithelium.
- To determine if cell fusion occurs between bone marrow and intestinal cells in acute colitis.
Main Methods:
- Gender-mismatched bone marrow transplants from male to female mice.
- Induction of acute colitis using dextran sodium sulphate.
- Development of a novel multi-signal fluorescent in situ hybridization and staining technique to identify donor cells.
Main Results:
- No Y chromosome-positive cells were found in the epithelial compartment without CD45 positivity.
- No evidence of cell fusion was observed through combined X and Y chromosome analysis.
- Increased CD45-positive stromal/lymphoid cells and myofibroblasts were noted, with few Y-positive endothelial cells.
Conclusions:
- Whole bone marrow transplantation does not result in intestinal epithelial engraftment in this colitis model.
- The developed methodology aids in analyzing cell phenotypes post-transplant and in chimerism/regenerative medicine studies.
