Human bone marrow-derived mesenchymal stem cell sheets and methods for their production

A method for producing a human bone marrow-derived mesenchymal stem cell sheet comprising one or more layers of confluent human bone marrow-derived mesenchymal stem cells (hBMSCs), the method comprising:

• a) culturing hBMSCs in culture solution on a temperature-responsive polymer which has been coated onto a substrate surface of a cell culture support, wherein the hBMSCs in culture solution are a human clonal bone marrow-derived mesenchymal stem cell line generated from a single cell, and wherein the temperature-responsive polymer has a lower critical solution temperature in water of 0-80° C.;
• b) adjusting the temperature of the culture solution to below the lower critical solution temperature, whereby the substrate surface is made hydrophilic and adhesion of the cell sheet to the surface is weakened;
• and
• c) detaching the cell sheet from the culture support.

A method for producing a human bone marrow-derived mesenchymal stem cell (hBMSC) sheet comprising one or more layers of confluent human bone marrow-derived mesenchymal stem cells (hBMSCs), the method comprising:

• a) culturing hBMSCs in culture solution on a temperature-responsive polymer which has been coated onto a substrate surface of a cell culture support, wherein the hBMSCs in culture solution are a human clonal bone marrow-derived mesenchymal stem cell line generated from a single cell, and wherein the temperature-responsive polymer has a lower critical solution temperature in water of 0-80° C.;
• b) adjusting the temperature of the culture solution to below the lower critical solution temperature, whereby the substrate surface is made hydrophilic and adhesion of the cell sheet to the surface is weakened;
• c) detaching the cell sheet from the culture support;
• and
• d) culturing the cell sheet in culture solution containing interferon gamma (IFN-γ) or basic fibroblast growth factor (bFGF).