人类牙乳头细胞的体外培养及细胞生物学性状研究

目的　体外分离培养人类牙乳头细胞,并对传代细胞的生物学特性进行研究。方法　选择3~4月胎龄的自然流产人胚胎,分离牙乳头,组织块法培养人类牙乳头细胞并鉴定。观察细胞的生长特性,经Ⅰ型胶原、纤维粘连蛋白和层粘连蛋白免疫细胞化学染色及细胞矿化诱导后的钙盐染色对细胞的生物学性状进行研究。结果　分离培养的人类牙乳头细胞在体外生长良好,细胞及分泌基质中的Ⅰ型胶原、纤维粘连蛋白和层粘连蛋白表达阳性。将培养细胞行矿化诱导后,细胞可形成钙化基质。结论　通过机械分离及贴壁法可获得人类牙乳头细胞,其传代细胞在基质形成和矿化能力方面与体内人牙乳头细胞有相似性,有潜力作为牙组织工程的种子细胞。

Culture and Characteristics of Human Dental Papilla Cells in vitro

OBJECTIVE: To culture human dental papilla cells (HDPCs)and to study its cytobiological characters in vitro. METHODS: HDPCs were isolated and cultured with explant culture technique in vitro; Type I collagen, fibronection and laminin were detected in HDPCs and its secreted matrix with the immunocyto-chemical stain; HDPCs were incubated in mineralized promoting solution containing 10 mmol/L beta-glycerophosphate, 100 mg/L of ascorbic acid and 10 nmol/L dexamethasone supplemented with 10% FBS and the form of mineralized nodules was tested with Alizarin Red S stainning. RESULTS: Cultured HDPCs in vitro were well growing in DMEM/F12. Type I collagen, fibronection and laminin staining were all positive in both HDPCs and its secreted matrix, and laminin was stained with bunchiness in matrix. Mineralized nodules formed after cultured 27 days by Alizarin Red S stainning. CONCLUSION: HDPCs isolated and cultured are well growing in vitro, have a capability of synthesizing and secreting matrix and in mineralized promoting solution, are able to form mineralizer, so, HDPCs have a capacity of seed cell of tissue engineering regeneration tooth.