Effect of rocking culture on keratinocyte differentiation in three-dimensional reconstitution human epidermis

A three-dimensional culture system of keratinocytes is able to mimic the formation of stratified epidermis. Air-exposure treatment is essential to promote keratinocyte differentiation. During the culture, we compared the transcripts in the cells with (Air) and without (Liq; submerged) air-exposure. Then, since rocking (shaking) the culture dish enabled to recover the keratinocyte differentiation despite the submerged culture (Liq), a comparative study between with and without rocking was also carried out to investigate the characteristics of the restored differentiation by RNA-seq. Transcriptome analyses revealed the expected similar pattern of the transcripts between air-exposure and rocking culture, which differed from those in Liq culture. Overall design: To investigate the biological significance, RNAs from three different culture systems (air-exposure, the submerged culture without air-exposure, rocking in the submerged cultures) were prepared at the same time. In each culture, RNA were prepared from three independent cultured cells and subjected to RNA-seq analysis. Comparative gene expression profiling analysis of RNA-seq data was carried out for each culture system.