Method for producing keratinocytes, method for producing cultures for epidermal evaluation, and culture substrate for keratinocytes

Abstract

It suppresses the aging of keratinocytes, promotes the differentiation of keratinocytes, and maintains the epidermal stem cells contained in the culture. [Solution] The method for producing keratinocytes includes the steps of culturing mesenchymal stem cells on a substrate, and culturing keratinocytes on a cell culture substrate obtained by decellularizing the cultured mesenchymal stem cells.

Claims

[Claim 1] The process includes culturing mesenchymal stem cells on a substrate, and culturing keratinocytes on a cell culture substrate obtained by decellularizing the cultured mesenchymal stem cells. A method for producing keratinocytes. [Claim 2] The method for producing keratinocytes according to claim 1, wherein the decellularization treatment includes at least one selected from immersion in a surfactant, immersion in an alkaline aqueous solution, repeated freeze-thaw cycles, and application of osmotic shock. [Claim 3] The method for producing keratinocytes according to claim 1, wherein the mesenchymal stem cells are at least one cell selected from the group consisting of primary bone marrow-derived mesenchymal stem cells, primary adipose-derived mesenchymal stem cells, mesenchymal stem cell lines, and mesenchymal stem cells obtained by differentiating pluripotent stem cells. [Claim 4] The method for producing keratinocytes according to claim 1, wherein the mesenchymal stem cells are cells that express at least one gene selected from the group consisting of fibronectin gene, type I collagen gene, versican gene, and biglycan gene. [Claim 5] The method for producing keratinocytes according to claim 1, wherein the mesenchymal stem cells have the ability to differentiate into one selected from the group consisting of osteoblasts, adipocytes, and chondrocytes. [Claim 6] The process involves culturing mesenchymal stem cells on a substrate, and then culturing keratinocytes on a cell culture substrate obtained by decellularizing the cultured mesenchymal stem cells. The process of obtaining a culture in which the keratinocytes have been differentiated, A method for producing a culture for epidermal evaluation containing the following. [Claim 7] The method for producing a culture for epidermal evaluation according to claim 6, wherein the decellularization treatment includes at least one selected from immersion in a surfactant, immersion in an alkaline aqueous solution, repeated freeze-thaw cycles, and application of osmotic shock. [Claim 8] The method for producing a culture for epidermal evaluation according to claim 6, wherein the mesenchymal stem cells are at least one cell selected from the group consisting of primary bone marrow-derived mesenchymal stem cells, primary adipose-derived mesenchymal stem cells, mesenchymal stem cell lines, and mesenchymal stem cells obtained by differentiating pluripotent stem cells. [Claim 9] The method for producing a culture for epidermal evaluation according to claim 6, wherein the mesenchymal stem cells are cells that express at least one gene selected from the group consisting of fibronectin gene, type I collagen gene, versican gene, and biglycan gene. [Claim 10] The method for producing a culture for epidermal evaluation according to claim 6, wherein the mesenchymal stem cells have the ability to differentiate into one selected from the group consisting of osteoblasts, adipocytes, and chondrocytes. [Claim 11] A culture substrate for keratinocytes obtained by culturing mesenchymal stem cells on a substrate and then decellularizing the cultured mesenchymal stem cells. [Claim 12] The keratinocyte culture substrate according to claim 11, wherein the decellularization treatment comprises at least one selected from immersion in a surfactant, immersion in an alkaline aqueous solution, repeated freeze-thaw cycles, and application of osmotic shock. [Claim 13] The keratinocyte culture substrate according to claim 11, wherein the mesenchymal stem cells are at least one cell selected from the group consisting of primary bone marrow-derived mesenchymal stem cells, primary adipose-derived mesenchymal stem cells, mesenchymal stem cell lines, and mesenchymal stem cells obtained by differentiating pluripotent stem cells. [Claim 14] The keratinocyte culture substrate according to claim 11, wherein the mesenchymal stem cells are cells that express at least one gene selected from the group consisting of fibronectin gene, type I collagen gene, versican gene, and biglycan gene. [Claim 15] The keratinocyte culture substrate according to claim 11, wherein the mesenchymal stem cells have the ability to differentiate into one selected from the group consisting of osteoblasts, adipocytes, and chondrocytes.

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