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Method for generating autologous retinal stem cells and autologous retinal cells by reversely differentiating human body cells, kit and application of autologous retinal stem cells and autologous retinal cells

A somatic cell and retinal technology, applied in the field of biomedicine, can solve the problems of long-term efficacy and safety that cannot be promoted clinically

Inactive Publication Date: 2012-07-25
BEIJING CSC CREATION BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the use of allogeneic retinal stem cells differentiated from human embryonic stem cells has successfully restored the vision of patients with retinitis pigmentosa; however, it cannot be clinically promoted due to long-term efficacy and safety issues

Method used

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  • Method for generating autologous retinal stem cells and autologous retinal cells by reversely differentiating human body cells, kit and application of autologous retinal stem cells and autologous retinal cells
  • Method for generating autologous retinal stem cells and autologous retinal cells by reversely differentiating human body cells, kit and application of autologous retinal stem cells and autologous retinal cells
  • Method for generating autologous retinal stem cells and autologous retinal cells by reversely differentiating human body cells, kit and application of autologous retinal stem cells and autologous retinal cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1: Production of plant and protein-induced retinal stem cells using peripheral venous blood / umbilical cord blood as raw material cells

[0074] Peripheral venous blood and umbilical cord blood (donated by scientific research staff, Beijing Wangjing Hospital, Beijing Wuzhou Women's Hospital, etc.) were collected aseptically. Before collecting peripheral venous blood / umbilical cord blood, the consent of the blood donor or his immediate family members should be obtained, and the genetic and infectious disease history of the blood donor and his family, as well as all relevant virus test results in the hospital should be recorded.

[0075] Peripheral venous blood / cord blood was routinely anticoagulated and stored at 4°C. Send to the stem cell production center within 24 hours. After the center needs to do the corresponding virus detection again, it will enter the computer registration procedure of the center, and after recording the corresponding barcode number, t...

Embodiment 2

[0078] Example 2 Using peripheral venous blood / umbilical cord blood as raw material cells to produce plant and protein-induced retinal cells

[0079] Peripheral venous blood and umbilical cord blood (donated by scientific research staff, Beijing Wangjing Hospital, Beijing Wuzhou Women's Hospital, etc.) were collected aseptically. Before collecting peripheral venous blood / umbilical cord blood, the consent of the blood donor or his immediate family members should be obtained, and the genetic and infectious disease history of the blood donor and his family, as well as all relevant virus test results in the hospital should be recorded.

[0080] Peripheral venous blood / cord blood was routinely anticoagulated and stored at 4°C. Send to the stem cell production center within 24 hours. After the center needs to do the corresponding virus detection again, it will enter the computer registration procedure of the center, and after recording the corresponding barcode number, the blood ...

Embodiment 3

[0090] In this example, the safety study of transplantation of plant and protein-induced autologous retinal stem cells prepared in Example 1 on nude mice is carried out as follows.

[0091] In two groups of nude mice transplantation experiments, nude mice were subcutaneously transplanted and injected with 1×10 8 plant and protein-induced retinal stem cells or 1×10 8 Personal natural embryonic stem cells (Beijing Wangjing Hospital), observe whether there is tumor formation at the inoculation site. Continuous observation for 90 days.

[0092] The results showed that none of the 10 nude mice in the plant and protein-induced retinal stem cell test group developed tumors, which was relatively safe; while all the 10 nude mice in the human natural embryonic stem cell test group developed tumors.

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Abstract

The invention provides a method for generating autologous retinal stem cells and autologous retinal cells by reversely differentiating human body cells. The method comprises the following steps of: culturing the human body cells in steps in a culture medium which contains different plant extracts and proteins, so that the human body cells are reversely differentiated to generate the autologous retinal stem cells and then continuously generate a plurality of autologous retinal cells. Tens of billions of a first generation of autologous retinal stem cells are generated within 2 to 3 weeks. The plant and protein-induced human autologous retinal stem cells have the same cell specific phenotype as human natural retinal stem cells and various cell differentiation abilities. The autologous retinal stem cells and the autologous retinal cells have good application prospect in the field of treating diseases, such as retinal pigment degeneration, and also have good potential application prospect in the fields of regenerative medicine and autologous tissue organ engineering. The autologous retinal stem cells and the autologous retinal cells have the possibility to be produced massively in an industrialization way.

Description

technical field [0001] The field of the present invention belongs to the technical field of biomedicine. Specifically, the present invention relates to a cell culture method and application for reverse differentiation of human somatic cells to produce autologous retinal stem cells and autologous retinal cells. Background technique [0002] With the advancement of medical science and the continuous improvement of diagnosis and treatment methods, infectious eye diseases and cataracts and other blinding eye diseases have been effectively diagnosed and treated. At the same time, the incidence and blinding rate of retinal degenerative diseases are on the rise. It is one of the major blinding eye diseases in the world. At present, its treatment is not only a hot spot and difficulty in ophthalmology but also neuroscience research. [0003] Although in the past ten years, some scholars at home and abroad have used retinal transplantation, gene therapy, drugs, cytokines, and artifici...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/074A61K35/12A61P27/02
CPCC12N5/062C12N2506/115A61P27/02
Inventor 林雄斌石海鹏李贵铃任海英廖焕昭
Owner BEIJING CSC CREATION BIOTECH
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