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Hou F.-Y.,Lanzhou University | Xie X.-W.,Pharmaceutical Research Institute of Gansu Province | Xi F.-Q.,Peoples Hospital of Guangxi Autonomous Region | Xu S.-H.,Lanzhou University | And 2 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2013

Objective: To study the effects of muscone-containing serum on proliferation and osteogenic differentiation of mesenchymal stem cells (BMSCs) to understand the action mechanism of muscone monomer on mesenchymal stem cells. Methods: Adherence screening was used to extract rat BMSCs, which were then cultured to the third generation. BMSC surface antigen as well as osteogenic and adipogenic induction were detected by flow cytometry. Then different concentrations of muscone-containing serum were used for intervention when successful culture was identified. Cell proliferation rate, ALP activity, calcium deposition, and osteocalcin secretion were determined to study the effects of muscone-containing serum on the proliferation and differentiation of rat stem cells. Results: Screened purified BMSCs showed homogeneous fibroblast-like, adherent growth, mainly with long fusiform. CD45 was negative while CD44 and CD90 were positive. The proliferation rate, ALP activity, calcium deposition, osteocalcin secretion, and calcified nodules were higher in muscone-containing serum intervention group than in the blank control group. Conclusion: Muscone-containing serum can promote the proliferation and osteogenic differentiation of BMSCs. Source


Xie X.-W.,Pharmaceutical Research Institute of Gansu Province | Hou F.-Y.,Lanzhou University | Li N.,Lanzhou University | Li S.-H.,Pharmaceutical Research Institute of Gansu Province | Song M.,Lanzhou University
Chinese Journal of Tissue Engineering Research | Year: 2013

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. Source

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