牙周膜成纤维细胞多种培养法的研究

Study on the various culture methods of periodontal ligament fibroblast cell

目的提高人牙周膜成纤维细胞原代培养的成功率,提高组织块贴壁率,缩短培养时间,加快细胞增殖。方法以高血清结合改良组织块培养法培养原代HPDLCs与传统组织块法及改良组织块培养法作比较。通过免疫组化方法鉴定细胞的来源,绘制细胞的生长曲线,对三种方式的培养成功率、组织块贴壁率及细胞增殖速度进行比较。结果三种方法所培养的原代细胞生长均趋于正常,细胞呈长梭形或多角形,波形蛋白鉴定呈阳性,角蛋白鉴定呈阴性,符合正常HPDLCs的正常形态及生物学特点。高血清结合改良组织块培养法的成功率为86.67%,明显高于其他两种方法(P<0.01)。结论高血清结合改良组织块培养法明显提高了HPDLCs原代培养的成功率,适合在临床上运用。

Objective To improve the successful rate of primary culture of periodontal fibroblast cells and tissue block adherence rate,reduce culture time and increase cell production.Methods Culture primary HPDLCs by combining high serum and improved tissue block,which was compared with traditional tissue block method and improved block method.Recognize the source of cell by immunity group,draw the growing curve of cell,and make comparison of successful rate,adherence rate and production rate of three methods.Results The primary cells cultured by three methods are all normal.The cells look like fusiform and multi-angle shape,vimentin positive,Keratin negative,which conform of normal morphology of HPDLCs and biological characteristics.The successful rate of improved tissue block with high serum totals 86.7%,obviously higher than that of other two groups（P〈0.01）.Conclusion The method of high serum improved tissue block obviously is increased the successful rate of primary cultivation,which shall be applied in clinical cases.