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Method for generating hiPSC-RPE cell sheet in combination with biological matrigel

A technology of cell membrane and biomatrix, which is applied in the field of combining biomatrix glue to generate hiPSC-RPE cell membrane, to achieve the effect of promoting clinical transformation and application, and good biocompatibility

Pending Publication Date: 2022-05-27
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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Problems solved by technology

[0003] The preparation of existing RPE sheets is mainly based on synthetic materials, which have many differences from the environment of natural BM membranes. There is no use of hiPSC-derived RPE to prove the safety and effectiveness of Uvea-Gel

Method used

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  • Method for generating hiPSC-RPE cell sheet in combination with biological matrigel
  • Method for generating hiPSC-RPE cell sheet in combination with biological matrigel
  • Method for generating hiPSC-RPE cell sheet in combination with biological matrigel

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Embodiment Construction

[0032] In order to make the purpose and advantages of the present invention clearer, the present invention will be further described below in conjunction with the embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention, and are not intended to limit the present invention.

[0033] Preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are only used to explain the technical principle of the present invention, and are not intended to limit the protection scope of the present invention.

[0034] It should be noted that, in the description of the present invention, the terms “upper”, “lower”, “left”, “right”, “inner”, “outer” and other terms indicated in the direction or the positional relationship are based on the drawings. The direction or positional relationship shown is onl...

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Abstract

The invention relates to a method for generating a hiPSC-RPE cell sheet by combining with biological matrigel, which comprises the following steps: step S1, preparing decellularized matrigel from pretreated pig eye grape membrane; step S2, maintaining the culture of the hiPSCs; step S3, culturing the hiPSC-RPE suspension ball; step S4, carrying out multiplication culture on the hiPSC-RPE cells; step S5, generating a hiPSC-RPE cell membrane sheet; wherein in the step S5, a Uvea-Gel coated culture dish with the concentration of 1% is placed at the temperature of 37 DEG C for about 30 min, and after Uvea-Gel is solidified, a layer of thin film sample area, namely a BM film, is formed and is used for simulating a natural BM microenvironment. According to the method, the decellularized matrix hydrogel separated and prepared from pig choroidal tissue is optimized, the hiPSC-RPE cells are obtained in combination with an established hiPSC-induced 3D retinal differentiation system, a method for generating the RPE sheet by simulating a Bruch membrane (BM) microenvironment is established, and a safe and effective biological material is provided for culturing GMP-level stem cells.

Description

technical field [0001] The invention relates to the field of hiPSC-RPE cell membranes, in particular to a method for generating hiPSC-RPE cell membranes by combining biological matrigel. Background technique [0002] At room temperature, Matrigel polymerizes to form a biologically active three-dimensional matrix, which simulates the structure, composition, physical properties and functions of the basement membrane of cells in vivo, which is beneficial to the culture and differentiation of cells in vitro, and can be used for cell morphology, biochemical function. , migration, infection and gene expression studies. At present, Matrigel has been extracted from EHS mouse tumors rich in extracellular matrix proteins. The main components are laminin, type IV collagen, nestin, heparin sulfate glycoprotein, growth factors and matrix metals protease etc. [0003] The preparation of existing RPE sheets is mainly based on synthetic materials, which are quite different from the enviro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12N5/074
CPCC12N5/0621C12N2533/90C12N2506/45C12N2513/00C12N2535/00
Inventor 李桂兰刘奕志郑颖丰
Owner ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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