三种组织块法培养人牙周膜细胞的比较研究

Comparative Study of Explant Culture of Human Periodontal Ligament Cells by Three Different Methods

目的提高人牙周膜细胞(Human periodontal ligament cells,HPDLCs)原代培养的成功率,并缩短培养周期。方法以高血清培养基组织块法、传统组织块法和玻片覆盖组织块法等3种方法原代培养HPDLCs。通过镜下观察细胞形态,免疫组化方法鉴定细胞来源,及绘制细胞生长曲线等方面,对各种方法的成功率进行比较。结果 3种方法所培养的原代细胞均生长良好,呈梭形,波形丝蛋白和碱性磷酸酶染色阳性、角蛋白染色阴性,符合HPDLCs的形态和生物学特征。高血清培养基组织块法的原代培养的成功率为84.21%,明显高于其他2种方法 (P<0.01)。结论高血清培养基组织块法能显著提高HPDLCs原代培养的成功率。

Objective To improve the success rate of primary culture of human periodontal ligament cells （HPDLCs）, and reduce its culture period. Methods Primary HPDLCs were obtained via three methods of explant culture： high-serum media method, conventional serum supplemented media method and coverslip method. Morphological features was observed by microscopy. Biological features were investigated by the growth curve. And immunocytochemical staining was used to characterize the cell lineage The success rates were compared between the groups. Results The spindle-shaped cells grew well in vitro. Vimentin and alkaline phosphatase were positive, while keratin was negative.Their morphological and biological characteristics were similar to those of typical HPDLCs. The success rate of primary culture was the highest in high-serum group （84.21%）. Conclusion High serum in culture media will greatly improve the success rate of primary culture of HPDLCs.