Skin functions as a protective barrier between us and the outside world, with the keratinocytes of the epidermis forming the outermost air-liquid interface barrier. The goal of the present study is to create high-resolution snapshots for each step of keratinocyte differentiation using single cell isolation techniques, to better understand the link between dysregulation of keratinocyte differentiation with common skin diseases such as atopic dermatitis. The uppermost living layer, called stratum granulosum (SG), undergo cell death program to form air-liquid barrier interface known as stratum corneum (SC) . Each cell in the SG might be exposed to a unique set of environmental stimuli, which would in turn contribute to the overall phenotype of the epidermis, as these cells eventually undergo cell death to form the final SC barrier. Employing single-cell based analyses to study such heterogenous SG cell populations, will thus reveal the detailed mechanisms by which the skin interacts with constant environmental stimuli. Although the respective layers of the skin are normally difficult to separate as a pure population of cells. At present, we are developing isolation methods of mouse and human ‘single’ epidermal cells (mice and human reconstituted epidermis) from each layer of the differentiated epidermis (SB, SS and SG, respectively) (arranged in the order of differentiation: Fig 1), using micro-pipette system. As a pre-requisite for the present project, we have already established a comprehensive single-cell transcriptome analysis platform. Furthemore, we are now establishing new methods for studying the epigenome (i.e. DNA methylation) in a single karatinocyte.
Single Cell Analysis of Epidermal Differentiation
RIKEN Center for Integrative Medical Sciences
Laboratory for Skin Homeostasis
Deputy Team Leader
RIKEN Center for Integrative Medical Sciences (IMS), 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa, 230-0045, Japan