Ⅰ型胶原酶消化组织块法体外培养人牙髓细胞

目的 建立利用Ⅰ型胶原酶消化组织块体外培养人牙髓细胞方法。方法 通过组织块Ⅰ型胶原酶消化法进行人牙髓细胞体外培养，免疫细胞化学法鉴定细胞组织来源，进行细胞传代，酶组织化学法对体外复层生长的人牙髓细胞爬片进行碱性磷酸酶组织化学染色，并检测其矿化能力。结果 采用Ⅰ型胶原酶消化组织块法体外培养的原代牙髓细胞，在第15～20d出现汇合，第4～6代牙髓细胞在连续培养后具有复层生长特性，并具有显著的碱性磷酸酶活性．阳性染色强弱部位呈区域性分布，连续培养的牙髓细胞可形成矿化结节。结论 Ⅰ型胶原酶消化组织块法可以很好地进行人牙髓细胞体外培养，可以用酶组织化学法研究牙髓细胞的生物学特性。

Study on Enzymatically Released Human Dental Pulp Cells with Collagenase Ⅰ in vitro

OBJECTIVE: To investigate the alkaline phosphatase activity of enzymatically released human dental pulp cells with collagenase I in vitro. METHODS: We cultivated human dental pulp cells from connective tissue explants digested with collagenase I. Immunocytochemical staining was performed for characterization. When the subcultured cells grew in multilayers, the activity of alkaline phosphatase was examined by enzyme histochemical staining,and the ability of mineralization was detected. RESULTS: Cultures of human primary dental pulp cells became confluent after 15-20 days. Subcultured cells proliferated to multilayers in long-term cultures, and the staining of alkaline phosphatase was noted to be regional on culture slides. The pulp cells cultured in the presence of beta-glycerophosphate and L-ascorbic acid formed mineralization nodules. CONCLUSION: These results suggest that human dental pulp cells can be cultivated preferably from tissue explants digested with collagenase I in vitro. Enzyme histochemistry is useful for studying the biological behavior of dental pulp cells.