Hair follicle epithelial stem cell preparation and hair follicle primordium preparation

Abstract

Epithelial stem cells (15), such as hair follicle epithelial stem cells, are cultured on a scaffold (13) comprising a molecule containing an extracellular cadherin (cadherin EC) domain. The epithelial stem cells 15 are then separated from the scaffold 13. The epithelial stem cells 15 are prepared through the abovementioned process. Thereafter, a hair follicle primordium10 may be produced by three-dimensionally culturing the epithelial stem cells 15 and mesenchymal cells 16 in a state in which a layer of the epithelial stem cells 15 and a layer of the mesenchymal cells 16 overlap. The mesenchymal cell 16 layer may include a layer of hair papilla cells and a layer of cells constituted of dermal pluripotent cells (DDMCs) or dermal sheath cells (DS cells). The epithelial stem cell 15 layer may overlap the hair papilla cell-layer side of the mesenchymal cell 16 layer.

Claims

1. A method for preparing hair follicle epithelial stem cells, comprising culturing hair follicle epithelial stem cells on a scaffold consisting of molecules containing a cadherin EC (Extracellular Cadherin) domain, and separating the hair follicle epithelial stem cells from the scaffold.

2. The method according to claim 1, wherein the scaffold is formed on the flat bottom surface of a container for cell culture.

3. The method according to claim 1, wherein the molecule is a cell-recognizing chimeric antibody E-cadherin Fc protein.

4. The method according to claim 1, wherein the hair follicle epithelial stem cells are proliferated by monolayer culture of the hair follicle epithelial stem cells on the scaffold.

5. The method according to claim 1, wherein, after culture, the hair follicle epithelial stem cells are CD34-negative, Itgb5-positive, and CD49f-positive.

6. A method for producing a hair follicle primordium, comprising preparing hair follicle epithelial stem cells by the method of any one of claims 1 to 5, and then culturing these cells in three dimensions with a layer of hair follicle epithelial stem cells and a layer of mesenchymal cells overlapping.

7. A method for producing a hair follicle primordium, comprising preparing hair follicle epithelial stem cells by the method of any one of claims 1 to 5, and then culturing these cells in three dimensions with a layer of hair follicle epithelial stem cells and a layer of dermal papilla cells overlapping.

8. A method for producing a hair follicle primordium, comprising: preparing hair follicle epithelial stem cells by the method of any one of claims 1 to 5; and then three-dimensionally culturing these cells with a layer of hair follicle epithelial stem cells and a layer of mesenchymal cells overlapping, wherein the layer of mesenchymal cells comprises a layer of dermal papilla cells and a layer of at least one of DDMCs (dermal pluripotent cells) and DS cells (dermal sheath cells), and the layer of hair follicle epithelial stem cells overlaps the layer of mesenchymal cells on the side of the layer of dermal papilla cells.

9. A method for producing a biopharmaceutical to be transplanted into the skin of a target, comprising: preparing hair follicle epithelial stem cells by the method of any one of claims 1 to 5; then producing a hair follicle primordium by three-dimensional culture of the cells in which a layer of hair follicle epithelial stem cells and a layer of mesenchymal cells overlap; and preparing a biopharmaceutical containing the hair follicle primordium, wherein the hair follicle epithelial stem cells and the mesenchymal cells are obtained by growing cells collected from the target.

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