Abstract: Objective: To observe and compare two methods to purify bone marrow mesenchymal stem cells (BMSCs) of green fluorescent protein(GFP) transgenic mice and study the feasibility to use green fluorescence protein(GFP)-BMSCs as a labeling method in transplantation experiment. Methods: Bone marrow cells isolated from 4 to 6 weeks old GFP transgenic mice were directly planted in culture flask. Cells were trypsinized on attaining 80% confluency in different Methods: (A)Regular method (B) Modified method. Morphological changes and intensity of the green fluorescent protein were observed under fluorescence microscope. Activity of BMSCs was identified according to the growth curve and multi-differentiation functions of BMSCs. Furthermore, the expression of some surface markers and green fluorescent protein was investigated by flow cytometry. Results: Because of the unwanted growth of hematopoietic cells and non-MSCs, lots of polymorphic cells were isolated from mouse via plastic adherent cultures. Compared with traditional way, the modified method could purify cells more efficiently. The subcultured cells maintained the capacity for amplification and were capable of differentiating into osteocytes and adipocytes. Their fluorescence was not diminished after several passages and could thus be used as a labeling method in vivo. Conclusion: The modified method is an effective method for purifying BMSCs of GFP transgenic mice.

Key words: GFP transgenic murine , Bone marrow mesenchymal stem cells, Modified method , Improve , Purification