| Objective To observe the labeling conditions of rat BMSCs using Hoechst33342, BrdU, and DAPI, respectively, and to compare the effect of each method. To observe cell growth and the fluorescence duration by culturing mouse BMSCs of transgenic green fluorescence mice, and to compared the superiority between mouse BMSCs and rat stem cells labeled with fluorescent. Methods Rat BMSCs were extracted using adherence screening, and cultured to the 3rd generation. The surface antigen was detected using flow cytometry. Staining was performed after osteogenic or adipogenic induction. Rat BMSCs were labeled with Hoechst 33342, BrdU, or DAPI, respectively. The proliferation of labeled rat BMSCs was detected. The fluorescence duration was observed under microscopy. Cell morphology and fluorescence duration of mouse BMSCs, which were cultured for different time, were observed. The advantages and disadvantages were compared with those of the 3 labeling methods mentioned above. Results Hoechst 33342, BrdU, and DAPI were available for BMSCs labeling. The labeling process of BrdU was long. None fluorescence quenching existed in the pre-labeled. Cells with positive labeling presented after the process of fluorescent immunohistochemistry. The labeling process of Hoechst 33342 and DAPI was short. The fluorescence duration was long. Because light should be avoided during the whole process, this brought some difficulties to the operation. No effect of the three methods on cell proliferation rate had been observed. The growth of BMSCs of transgenic mice was much slower than that of rat BMSCs. Its advantages were free of labeling and long fluorescence duration. So it was more convenient and feasible for cell tracking study. Conclusion Stem cells have the proficiency of multiple differentiations. They have great value of research and application. Stem cell labeling can provide a better platform for the mechanism study of stem cell migration and homing. |