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Method for inducing human embryonic stem cells to differentiate into retinal pigment epithelial cells in vitro

A technology for human embryonic stem cells and retinal pigment, applied in the biological field, can solve the problems of large difference between batches, low cell yield, and complicated methods, and achieve the effects of simple method, high cell rate and long differentiation time.

Active Publication Date: 2020-04-07
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, these reported methods have their own advantages and disadvantages, such as long time-consuming, cumbersome methods, large batch-to-batch variance and low cell yield still exist.

Method used

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  • Method for inducing human embryonic stem cells to differentiate into retinal pigment epithelial cells in vitro
  • Method for inducing human embryonic stem cells to differentiate into retinal pigment epithelial cells in vitro
  • Method for inducing human embryonic stem cells to differentiate into retinal pigment epithelial cells in vitro

Examples

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Embodiment

[0027] This example is a method for efficiently inducing hESCs to differentiate into RPE cells in vitro.

[0028] In this embodiment, the induction of differentiation includes the following steps:

[0029] a. Inoculate hESCs into feeder cells, add complete medium for stem cells, place for culture and change the medium regularly until the cells reach confluence;

[0030] The hESCs described in this step is the cell line H1, and the feeder layer cells are X-ray treated mouse embryonic fibroblasts; the components of the complete medium for stem cells are: DMEM / F12 basal medium, serum substitute, non-essential amino acids, penicillin , streptomycin, glutamine, β-mercaptoethanol and fibroblast growth factor, wherein the volume content of serum replacement is 20%, the concentration of non-essential amino acid is 0.01mmol / L, and the concentration of penicillin is 100U / ml , the concentration of streptomycin was 100ug / ml, the concentration of glutamine was 1mmol / L, the concentration o...

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Abstract

The invention relates to a method for inducing human embryonic stem cells to be differentiated into retinal pigment epitheliums (RPE) in vitro, and belongs to the biology technical field. The pure RPEs can be obtained through one-time passage by means of induced differentiation. Compared with the prior art, the differentiation time from the human embryonic stem cells into the RPEs is shortened. By the adoption of the method, a new approach and guidance are provided for induced directional differentiation of the human embryonic stem cells into the pigment epitheliums.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing human embryonic stem cells to differentiate into retinal pigment epithelial cells in vitro. Background technique [0002] Degenerative retinal disease is the leading cause of blindness worldwide. In this disease, damage and dysfunction of retinal pigment epithelium (RPE) lead to degeneration of photoreceptor cells, eventually affecting the patient's vision until blindness. Due to the non-regeneration of RPE and photoreceptor cells, there is no effective treatment for this kind of disease in clinical practice. Cell therapy holds more promise for the treatment of retinal degenerative diseases than any other treatment. [0003] Because embryonic stem cells (ESCs) have the ability to proliferate indefinitely and differentiate into various cells, they can be used as an ideal source of transplanted cells. Reports have confirmed the feasibility and safety of ESCs-de...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 徐国彤高芙蓉李宗义吕立夏
Owner TONGJI UNIV
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