Effect of CM-Dil labeling on biological characteristics of rat bone marrow mesenchymal stem cells

Zhang Ya-ni; Yu Mei-juan; Feng Shan-wei; Wang Shu-hui; Li Mei-shan; Xiong Fu; Zhang Cheng

doi:10.3969/j.issn.2095-4344.2016.45.006

Abstract：BACKGROUND:Existing evidence has shown that CM-Dil-labeled mesenchymal stem cel s have delivered better results in experiments in vivo and in vitro. <br> OBJECTIVE:To investigate the effect of in vitro CM-Dil labeling on biological characteristics and directed migration of rat bone marrow mesenchymal stem cel s. <br> METHODS:Mesenchymal stem cel s were isolated and cultured from the bone marrow of Sprague-Dawley rats. The specific surface antigens, CD29, CD44, CD11b, CD45, were detected by flow cytometry. The rat bone marrow mesenchymal stem cel s were labeled with CM-Dil, and then were cultured and passed continual y. Thereafter the morphology and proliferation ability of labeled cel s were assessed. The strength and duration of fluorescence after CM-Dil labeling were detected. Transwel chambers were used to investigate the migration ability of labeled cel s towards the homogenate of the gastrocnemius muscles of mdx mice. <br> RESULTS AND CONCLUSION:The cultured cel s were markedly positive for CD29 and CD44, but negative for CD11b and CD45, which were in accordance with the features of bone marrow mesenchymal stem cel s. CM-Dil-labeled cel s were not different from unlabeled cel s in the aspect of morphology and proliferation. Fluorescence was stil visible after passage;however, a reduction in fluorescent intensity was observed under an inverted fluorescent contrast phase microscope after labeling. The labeled cel s showed no different migration ability from the unlabeled cel s (P>0.05). In conclusion, the procedure of CM-Dil labeling is safe, simple and effective. CM-Dil could be a good choice for labeling and tracing rat bone marrow mesenchymal stem cel s.