Selecting keratinocytes for producing three-dimensional epidermal models, involves providing lysate from keratinocytes of epidermal sample, determining gene expression of four different genes from cell lysate, and selecting keratinocytes

摘要

Selecting keratinocytes for producing three-dimensional epidermal models, involves: (a) providing a lysate from the keratinocytes of an epidermal sample; (b) determining the gene expression of four different genes from the cell lysate; (c) determining the gene expression of identical genes in the cell lysate of a reference sample; and (d) selecting the keratinocytes, in which the gene expression of at least three genes is greater than the gene expression of identical genes and in which the gene expression of at least one gene is smaller than the gene expression of the same gene. Selecting keratinocytes for producing three-dimensional epidermal models, involves: (a) providing a lysate from the keratinocytes of an epidermal sample; (b) determining the gene expression of four different genes from the cell lysate provided in the step (a), where the four different genes are (i) genes encoding proteins and/or enzymes of the terminal differentiation of keratinocytes, (ii) genes encoding proteins and/or enzymes of lipid metabolism of keratinocytes, (iii) genes encoding proteins and/or enzymes for the structure of a skin barrier, (iv) genes encoding proteins of cell-cell adhesion, (v) genes encoding proteins that occur in inflammatory processes of the skin, (vi) genes encoding regulatory proteins that affect the metabolism of the skin, and (vii) regulatory RNA molecules; (c) determining the gene expression of identical genes as in the step (b) in the cell lysate of a reference sample, where the keratinocytes are used to prepare the cell lysate of the reference sample; and (d) selecting the keratinocytes, in which the gene expression of at least three genes determined in the step (b) is greater than the gene expression of identical genes determined in the step (c) and in which the gene expression of at least one gene determined in the step (b) is smaller than the gene expression of the same gene determined in the step (c). An independent claim is included for a kit comprising a reference sample, primers for four different genes, and optionally a mixture comprising at least one reverse transcriptase, nucleotide triphosphates, and optionally a ribonuclease inhibitor.