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| ZHONG Zhaoming,ZHANG Yu,ZHA Dingsheng.In vitro culture of adult human ligamentum flavum cells[J].Chinese Journal of Spine and Spinal Cord,2010,20(1):62-65. |
| In vitro culture of adult human ligamentum flavum cells |
| Received:October 09, 2009 Revised:November 04, 2009 |
| English Keywords:Ligament flavum Cell culture Degeneration |
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| English Abstract: |
| 【Abstract】 Objective：To explore the culturing of human ligamentum flavum（HLF） cells in vitro.Method：HLF were harvested during surgery for thoracolumbar burst fractures.HLF cells were isolated by collagenase-predigested explants cultures.Under an inverted phase microscope，cells were examined for outgrowth，morphology and growth status.After 1-8 days′ culturing，for HLF cells at the first，third and fifth passage（P1，P3，P5），the 3-（4，5-dimethy thioazol-2-yl）-2，5-di-phenytetrazoliumromide（MTT） method was used to analyze the cell proliferation by measuring the optical density（OD） values，and cell growth curves were drawed.Expressions of vimentin and type Ⅰ collagen in cells of third passage were detected by immunofluorescence staining.Result：Within 10-14 days after explants，outgrowth of cells was observed from ligament tissue explants and became monolayer.Morphologically，HLF cell lines varied widely in appearance，and arranged mainly as spindle-shape and polygonal-shape.At subconfluence，cells showed vortex-like growth.S-shaped growth curve was shown in the P1，P3 and P5 cells.The OD values at different time points showed significant difference compared with the same passage cells（P<0.001）.Significant differences in OD value was not found among the 3 different passage cells（P＞0.05）.No interaction effect was observed between cell passages and time points，the cell proliferation of the P1，P3 and P5 showed no significant difference（F=0.283，P=0.957）.Immunofluorescence staining showed expression of vimentin and type Ⅰ collagen.Conclusion：HLF cells can be successfully isolated by collagenase-predigested explant cultures.HLF cells in primary culture shows fibroblast-like phenotype.Biological characteristics of cells within 5 generations are stable. |
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