气-液面培养对角质形成细胞分化的影响

目的 研制角质形成细胞-胶原-猪去细胞真皮基质(PADM)组织工程皮肤,观察气-液面培养对角质形成细胞分化及细胞因子基因表达的影响.方法 将角质形成细胞种植在真皮基质胶原面进行气-液面培养和浸没式培养.采用逆转录-聚合酶链反应(RT-PCR)技术,对比研究两种培养方法获得的细胞膜片bFGF基因表达差异.结果 气-液面培养获得的角质形成细胞层结构与正常皮肤相似,细胞分化良好.单纯浸没式培养角质形成细胞层较薄,分化较差.气一液面培养获得的细胞膜片bFGF mRNA表达量明显高于单纯浸没式培养(t=3.825,P<0.01).结论 气-液面培养可促进角质形成细胞的生长、分化及复层表皮结构形成,影响细胞因子的基因表达.

The influence of air-liquid interface culture on differentiation of keratinocytes

Objective To prepare tissue-engineered skin containing keratinocytes and dermal matrix prepared by Coagen-PADM ,and observe the influence of air-liqnid interface culture on differentiation and cytokine gene expression of keratinocytes. Methods The keratinocytes were inoculated on the collagen surface of the dermal matrix by air-liquid interface culture and imergecl culture. By using RT-PCR method,the bFGF gene expression in keratinocytes sheets was compared. Results The structure of keratinocytes by air-liqnid interface culture was similar to that of the normal skin, and keratinocytes differentiated well. By using imerged culture method, the structure of keratinocytes was thin, and keratinocyte differentiation was immature. The UFGF-RNA expression of keratinocytes sheet was higher by air-liquid interface culture (t = 3.825 ,P<0.01). Conclusion Air-liquid interface culture may facilitate the growth, differentiation and multiple stratum of keratinocytes, and influence the cytokine gene expression.