酶消化组织块法原代培养人牙周膜成纤维细胞的初步研究

PRIMARY CULTURE OF HUMAN PERIODONTAL LIGAMENT FIBROBLASTS BY EXPLANTS WITH ENZYMATIC DIGESTION

目的 提高人牙周膜成纤维细胞 (humanperiodontalligamentfibroblasts,HPLF)原代培养的成功率 ,快速建立体外细胞研究模型。方法 采取组织块与酶消化相结合的方法处理牙周膜组织块 ,获取细胞 ;用形态学、免疫细胞化学染色等方法鉴定细胞来源 ,通过生长曲线的测定研究体外细胞生长特性。结果 原代培养成功率为 77 8%。获得的细胞呈梭形 ,抗波形丝蛋白染色呈阳性 ,抗角蛋白染色呈阴性 ,符合人牙周膜成纤维细胞的形态学特征和生物学特性 ,生长良好。结论 酶消化组织块法可显著提高人牙周膜成纤维细胞原代培养成功率 ,方法可行。

OBJECTIVE TO INCREASE THE SUCCESS RATE OF PRIMARY CULTURE OF HUMAN PERIODONTAL LIGAMENT FIBROBLASTS (HPLF), AND TO ESTABLISH AN EXPERIMENTAL MODEL FOR STUDYING HPLF IN VITRO.METHODS THE PRIMARY CELLS WERE ISOLATED FROM HUMAN PERIODONTAL LIGAMENT BY EXPLANTS WITH ENZYMATIC DIGESTION METHOD. MORPHOLOGICAL ANALYSIS AND IMMUNOCYTOCHEMICAL STAINING WERE USED TO CHARACTERIZE THE CELL LINEAGE, AND GROWTH CURVE ASSAY TO EVALUATE THE BIOLOGICAL FEATURES OF HPLF.RESULTS THE SUCCESS RATE OF PRIMARY CULTURE OF HPLF WAS 77.8%. CULTURED CELLS WERE SPINDLE-SHAPED, AND HAD A POSITIVE REACTION TO ANTIBODIES AGAINST VIMENTIN, AND A NEGATIVE REACTION TO ANTIBODIES AGAINST KERATIN. THEIR MORPHOLOGICAL AND BIOLOGICAL CHARACTERISTICS WERE SIMILAR TO THOSE OF TYPICAL HPLF. GROWTH OF HPLF OBTAINED BY THIS METHOD WAS SATISFACTORY.CONCLUSION THE SUCCESS RATE OF PRIMARY CULTURE OF HPLF IS SIGNIFICENTLY INCREASED BY EXPLANTS COMBINED WITH ENZYMATIC DIGESTION. THE METHOD IS SIMPLE AND FEASIBLE.