人胚成釉细胞体外培养的实验研究

The experimental study of cell culture of human ameloblast

目的 :探索成釉细胞离体培养方法 ,建立成釉细胞体外模型。方法 :运用组织细胞培养技术分离人胚成釉细胞进行原代培养 ,倒置显微镜下观察培养细胞生长特点 ,免疫组织化学染色检测培养细胞釉原蛋白。结果 :培养细胞成片生长 ,细胞形态结构清楚。免疫组化染色 ,多数细胞抗釉原蛋白抗体染色阳性。结论 :用消化培养法 ,胶原做基质饲养层 ,含 5 %小牛血清的HAMF -12作培养基并加入EGF、INS、HC 。

Objective:The main idea of this study was to explore the method of the normal ameloblast culture in vitro. Method:In experiments we chose human embryo enamel organs to digest them into single cell with digestion zyme. Primary culture was performed on collagen-coated culture plates.Result: The ameloblasts of human embryo were primarily cultured on collagen-coated culture dish having a typical epithelium cell characteristics. The culture cells start to adhere to the plates within 24 hours. Most of the culture cells adhere to the plates on the third days. They can connect tightly. the number of culture cells did not increase for the first 3～5 days , and then increased gradually. Expression of amelogenin was observed.Conclusion: Human ameloblast could be cultured for a longer time maitainning it's characteristics when HAMF-12 which is added EGF,INS,HC and 5% calf serum was used as culture media and collagen was used as substrase.