诱导表皮角质形成细胞体外分层的途径及机制

Induction in the stratification of epidermal keratinocyte in vitro

目的 探讨诱导表皮角质形成细胞体外分层途径及机制。方法 在体外培养状态下,利用气液界面培养法及高浓度钙离子诱导胎鼠皮片、表皮角质形成细胞分层,借助冰冻切片、免疫荧光及激光共聚焦等技术检测其分层层数及分化程度。结果 胚胎龄16d的胎鼠皮片经气液界面培养3d后,其表皮基底上角质形成细胞由2层增加为5层;经气液界面培养7d后,胎鼠皮片表皮基底上角质形成细胞的局部区域分层已可达8层,细胞呈现更为分化的形态。在高浓度Ca2 + (1 5mmol L)条件下培养7d后,培养角质形成细胞发生了分层,分化标记物角蛋白K10在分层角质形成细胞中的表达明显增加。结论 气液界面培养法及高浓度钙离子能诱导表皮角质形成细胞的体外分层,同时促进其分化。

Objective To explore the pathway and mechanism of the induction in the stratification of epidermal keratinocyte in vitro. Methods The stratification and differentiation of the cultured free skin graft from fetal mouse and the epidermal keratinocyte were induced and investigated, using air-liquid culture and high concentration calcium combined with the frozen section, the immunofluorescence as well as the confocal laser scanning. Results After being maintained at the interface of air-liquid for 3 d, the suprabasal layers of the fetal skin epidermis increased from three to five. When the maintenance at the interface of air-liquid prolonged to 7 d, the suprabasal layers of the fetal skin epidermis partially increased to eight, with more differentiated form. After being cultured in high Ca 2+ (1.5 mmol/L) medium for 7 d, the stratification of epidermal keratinocyte formed, in which the differentiation marker, keratin 10 expressed strongly. Conclusion The air-liquid culture and high-concentration calcium could induce not only the stratification but also the differentiation of the epidermal keratinocyte in vitro.