Preparation method of stent-free retinal pigment epithelial cell sheet

Abstract

The invention provides a method for differentiating pluripotent stem cells into retinal pigment epithelial cells through three-dimensional suspension culture, and belongs to the technical field of biotechnology. The method comprises the following steps: digesting and resuspending human induced pluripotent stem cells to obtain monomeric cells; inoculating a U / V type low-adhesion culture material with the monomeric cells, and sequentially carrying out suspension culture with a retinal nerve progenitor cell culture solution, an RPE cell differentiation induction solution and an RPE cell maturation differentiation solution to obtain hollow cell spheres formed by monolayer retinal pigment epithelial cells; the obtained hollow cell spheres are cut open and flatly laid, and the stent-free retinal pigment epithelium cell sheet can be obtained. The RPE cell sheet obtained by the method has curvature, carries basal membrane tissues differentiated together with RPE cells, does not need an exogenous stent material when being applied to clinical transplantation in the future, and immunological rejection can avoided. The method disclosed by the invention is good in operability, simple in induction process, high in repeatability and convenient for future clinical precise application.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, in particular to a method for preparing a scaffold-free retinal pigment epithelial cell sheet. Background technique [0002] The retina is a layer of transparent film structure located inside the eyeball wall, which is located on the inner surface of the choroid, and can be divided into 10 layers, from outside to inside are retinal pigment epithelium, photosensitive layer, outer boundary membrane, outer nuclear layer, outer plexus layer, inner nuclear layer, inner plexiform layer, ganglion cell layer, nerve fiber layer, and inner limiting membrane. Among them, the retinal pigment epithelium (RPE) is a layer of regularly arranged polygonal melanocytes close to the retinal nerve cells. RPE cells cannot regenerate. Once RPE cells die due to disease or injury, they can only be treated by cell transplantation. However, the source of donor RPE cells is very limited, and the transplantation of R...

AI Technical Summary

Problems solved by technology

This method induces pluripotent stem cells to differentiate into pigment epithelial cells through three-dimensional suspension culture retinal organoid technology, which provides a new source of autologous transplantable RPE cell sheets, and the RPE cell sheets do not require exogenous scaffold materials, which improves clinical RPE. The safety and effectiveness of cell transplantation solves the safety hazards caused by the transplantation of RPE cell sheets formed by scaffold materials in the prior art, and provides a new technical solution for the use of RPE cell sheet transplantation to treat RPE-deficient eye diseases

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