Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (49): 9146-9151.doi: 10.3969/j.issn.2095-4344.2012.49.005

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Hoechst33342 for labeling rat bone marrow mesenchymal stem cells

Xie Xing-wen1, Hou Fei-yi2, Li Ning2, Li Sheng-hua1, Song Min2   

  1. 1Pharmaceutical Research Institute of Gansu Province, Lanzhou 730050, Gansu Province, China; 2Gansu University of Traditional Chinese Medicine, Lanzhou 730000, Gansu Province, China
  • Received:2012-01-10 Revised:2012-03-19 Online:2012-12-02 Published:2013-01-16
  • About author:Xie Xing-wen☆, Doctor, Associate chief physician, Master’s supervisor, Pharmaceutical Research Institute of Gansu Province, Lanzhou 730050, Gansu Province, China xxw19726@hotmail.com
  • Supported by:

    Supported by: the National Natural Science Foundation of China, No.81060299*

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Abstract:

BACKGROUND: Hoechst33342 can be used for labeling bone marrow mesenchymal stem cells, but there are few reports on the optimal concentration of marker.
OBJECTIVE: To investigate the effect of different concentrations of Hoechst33342 on the proliferation and labeling rate of rat bone marrow mesenchymal stem cells and to investigate the duration time of fluorescence.
METHODS: Adherence screening method was used to extract rat bone marrow mesenchymal stem cells, and the cells were cultured to the third generation, then surface antigen was detected by flow cytometry, and stained after osteogenic and adipogenic induction. The proliferation rate and labeling rate of rat bone marrow mesenchymal stem cells were detected after treatment with different concentrations of Hoechst33342, and then the duration time was detected through the use of fluorescence microscope.
RESULTS AND CONCLUSION: Hoechst33342 can be used for labeling bone marrow mesenchymal stem cells, and the optimal concentration of Hoechst33342 was 5 mg/L, the duration time was long and there was no quenching within 30 days. Hoechst33342 damaged the cells after fluorescence excitation, and when the concentrations exceed10 mg/L, the marker could cause cell death. When the concentration was 7.5 mg/L, cell proliferation was inhibited; when the concentration was 2.5 mg/L, the duration for labeling was shorter, the fluorescence decreased at 7 days and disappeared at 14 days. Hoechst33342 can be used for labeling bone marrow mesenchymal stem cells, and the optimal concentration of the Hoechst33342 is 5 mg/L; phenotype identification shows the negative expression of CD45 and positive expression of CD44 and CD90.

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