Original articleCharacterization of rhodamine 123 low staining cells and their dynamic changes during the injured-repaired progress induced by 5-FU

ObjectiveTo investigate the characterization of intestinal epithelial stem cells stained by Rhodamine 123 (Rho) and analyze the dynamic changes of intestinal epithelial stem cells during the injured-repaired progress induced by 5-FU.MethodsMucosal cells were obtained from adult C57BL/6J mice. The Rho stained cells were sorted using FACS. The mouse model of intestinal mucosal injured-repaired was established by injecting 5-FU and sacrificed at different time post-injection, and the middle intestines were used for detecting the percentage of Rho low staining cell fraction by FACS and detecting the expression of the intestinal epithelial stem cells marker-musashi-1 (msi-1) by RT-PCR and immunohistochemistry.ResultsThe Rho stained intestinal mucosal cells were divided into three fractions: Rho low staining fraction (12.35%), Rho middle staining fraction (35.5%) and Rho strong staining fraction (50.5%). The cells in Rho low staining fraction expressed rich msi-1 and most of which were in the G0/G1 phase of cell cycle. After treatment of 5-FU, the intestinal mucous were damaged, although the number of msi-1 positive cells has a little decrease, there was no statistical difference among the mice at different time after injection (P > 0.05). However, the percentage of msi-1 positive cells increased significantly after injection (P < 0.01), and the percentage of msi-1 positive cells decreased gradually during the repaired procedure of the intestinal mucous. There was significant positive correction between the percentage of msi-1 positive cells and the percentage of Rho low staining cell fraction (r = 0.867, p < 0.01) after 5-FU injection.ConclusionsThe Rho low staining cell fraction from intestinal mucous contained rich intestinal epithelial stem cells, and the intestinal epithelial stem cell which expressed msi-1 played a key role in repairing the damage of intestinal mucous induced by 5-FU.