Abstract:

BACKGROUND: In vitro culturing, amplification and labeling are important links to harvest labeled rat bone marrow mesenchymal stem cells (BMSCs) that are satisfactory to animal experiment.
OBJECTIVE: To explore a convenient and practical method for separating and labeling BMSCs using whole bone marrow adherent culture, and labeling with PKH26 in vitro.
METHODS: BMSCs were isolated and cultivated from the bone marrow of rats by whole bone marrow adherent culture. Further purification was achieved by expansion at serial passages. The passage 3 BMSCs were cultured and labeled with PKH26. The growth, fluorescence intensity and serial subcuhivation of labeled BMSCs were analyzed with fluorescence microscope. The proliferation ability of these labeled cells was tested by MTT.
RESULTS AND CONCLUSION: BMSCs were isolated and purified successfully and effectively by the method of whole bone marrow adherent culture. The labeled BMSCs appeared red fluorescence. After 3 passages of serial subcultivation, the fluorescence intensity and the labeling rate of BMSCs were gradually decreased. The biological features such as morphology, growth in vitro were not affected by labeling. BMSCs can be successfully cultivated by whole bone marrow adherence method conveniently. Labeling the BMSCs with PKH26 is an effective and practical method.