| Objective: The dental pulp cells of rabbit were isolated and cultured in vitro, to studythe proliferation characteristics and to identify the cells surface marker. Methods: Thedental pulp cells of rabbit were cultured by enzymatic digestion method. the dental pulppassage cells of rabbit were cultured by limiting dilution clone technology to observe themorphological changes on primary cells and passage cells. calculate the cell survival ratefrom the third generations to the sixth generations. test the cell clone forming rate with thesecond generation. measure the cell growth curve from the second generation to the tenthgeneration. measure the cell proliferation cycle from the second generation to the tenthgeneration by flow cytometry to identify the cells surface marker: vimentin、CD44、osteonectin、DSP in second generation.Result: The cells cultured by enzymatic digestionmethod and limiting dilution clone technology were no difference on morphologicalchanges. the proportion of the cell survival rate was94.7%、95.8%、95.2%、95.3%.the cellsclone forming rate was8-26/2000.the proliferation rate was difference between differentgenerations of cells. there was no difference between the second、fourth、sixth generationsof cell by LSD-t detection. there was significant difference between second、fourth、sixthand eighth、tenth as the cells passaged, the proportion of S stage increased. There was nodifference between the second、fourth、sixth generations of cell by LSD-t detection. therewas significant difference between second、fourth、sixth and eighth、tenth. The cellssurface marker of dental pulp stem cells showed the positive expression,its concludevimentin、CD44、osteonectin、DSP. Conclusion: The rabbit dental pulp cells caneffectively proliferated in vitro. Immunochemistry staining showed the positive expressionof vimentin、CD44、osteonectin and DSP. it indicates that the dental pulp stem cells can beisolated from the rabbit dental pulp cells. It provide the new types of seed cells for dentaltissue engineering. |
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