人牙胚体外器官培养模型的建立

Establishment of human tooth germ culture model in vitro

目的 :建立人牙胚体外器官培养模型 .方法 :分泌前期人牙胚采用Trowell型支架培养法 ,RPMI 16 4 0培养基(含 2 0 0 g·L-1FBS ,2 0 0mg·L-1谷氨酰胺 ,2 0 0mg·L-1L 抗坏血酸 ,1× 10 -7mol·L-1维甲酸 ,10 0kU·L-1青霉素 ,10 0kU·L-1链霉素 ) ,在 37℃ ,5 0mL·L-1CO2 +95 0mL·L-1空气条件下进行培养 ,每 4 8h更换一次培养基 .体外培养 .2 ,4 ,6和 8d随机抽取牙胚进行组织学观察 .结果 :培养的牙胚组织结构关系与对照组一致 ,牙尖部形态进一步突出 ,细胞形态典型 .在 8d时 ,牙尖之间部分造釉器细胞和成牙本质细胞开始出现坏死 .结论

AIM: To establish an organ culture model of human tooth germ in vitro . METHODS: Trowell type bracket method was used. The presecretory stage human tooth germ was cultured in vitro in RPMI 1640 medium at 37℃, 50 mL·L -1 CO 2+950 mL·L -1 air condition for 8 days. The medium contained 200 g·L -1 FBS, 200 mg·L -1 glutamine, 200 mg·L -1 L ascorbic acid, 1×10 -7 mol·L -1 retinoic acid, 100 kU·L -1 penicillin and 100 kU·L -1 streptomycin. The culture medium was replaced every 48 h. The tooth germ was randomly selected for histological observation at day 2, 4, 6 and 8. RESULTS: Status of the cultured tooth germs was similar to that of controls. The shape of cusp became more distinct. The cells of tooth germ were typical. On day 8, some cells between the cusps began to necrose. CONCLUSION: A practical model for studying the normal development of human tooth germ cells and the effects of various factors acting on human tooth germ development is provided.