Method of preparing decellularized soft tissue, graft and culture material
It is intended to provide a method of preparing a decellularized soft tissue whereby swelling after an ultrahigh pressure treatment can be inhibited, etc. The above-described method of preparing a decellularized soft tissue comprises the application step wherein an ultrahigh hydrostatic pressure is applied to an isolated soft tissue in a medium to thereby disrupt cells in the soft tissue, and the removal step wherein the disrupted cells are removed. As the medium, use is made of an aqueous solution containing a water-soluble polysaccharide such as dextran.
1. A method of preparing a decellularized soft tissue in which soft tissue of animal origin is decellularized, the method comprising:
an application step of disrupting cells in the soft tissue by applying an ultrahigh hydrostatic pressure to an isolated soft tissue in a medium; and
a removal step of removing the disrupted cells,
wherein the medium is an aqueous solution containing a water-soluble polysaccharide;
wherein the ultrahigh hydrostatic pressure is at least 1000 atmospheres,
wherein the application step comprises a pressure limiting step of limiting the rate of pressure increase or pressure decrease in the applied pressure applied to the medium to inhibit solidification of the medium and damage to the structure of the decellularized tissue during the process of the pressure increase or pressure decrease by preventing the applied pressure applied to the medium from taking a value greater than or equal to a melt pressure of the medium that is calculated in advance based on the composition of the medium.
2. The method according to claim 1 , wherein the water-soluble polysaccharide is at least one type of polysaccharide selected from a group consisting of dextran, alginic acid, hyaluronic acid, trehalose, 2-methacryloyloxyethyl phosphorylchloine, and polyvinylpyrrolidone.
3. The method according to claim 1 , wherein the application step includes a temperature maintenance step of maintaining a temperature of the medium to be greater than or equal to a melting point of the medium at the ultrahigh hydrostatic pressure.
4. The method according to claim 1 , wherein the soft tissue is corneal tissue.