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Meaning of relative gene expression in multilayered cultures of epidermal keratinocytes
Author(s) -
Malaisse Jérémy,
Hermant Maryse,
Hayez Aurélie,
Poumay Yves,
Lambert de Rouvroit Catherine
Publication year - 2014
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/exd.12501
Subject(s) - involucrin , filaggrin , gene expression , biology , epidermis (zoology) , cellular differentiation , morphogenesis , microbiology and biotechnology , gene , regulation of gene expression , loricrin , genetics , immunology , anatomy , atopic dermatitis
Reconstructed human epidermis (RHE) has become an in vitro model of choice for studying cell and tissue functions. Analysis of gene expression over the course of reconstruction must take into account the heterogeneous differentiation states of keratinocytes reconstituting the typical epidermal layers. In monolayer cultures, relative mRNA expression levels of differentiation markers are usually expressed as a ratio versus a classical reference gene (also named house‐keeping gene) tested to be expressed equally in certain experimental conditions. Applied to complex tissues in which the cell number increases over time together with differentiation, calculation of relative gene expression does not take enough into account a crucial phenomenon: epidermal morphogenesis results in progressive restriction of differentiation markers, such as involucrin, to a specific layer, or in the delayed onset of mRNA expression of filaggrin or TMEM45A for instance following stratification. Our study illustrates that comparing the relative expression level of mRNA s to that of a basal layer‐specific gene (e.g. ITGA6) better illustrates the contribution of specific differentiation markers to the process of epidermal morphogenesis.