Induced pluripotent stem cell line for reducing cell immunogenicity and construction method thereof

A technology of pluripotent stem cells and cellular immunity, applied in the field of induced pluripotent stem cell lines knocking out human leukocyte antigen-related genes, can solve the problems of difficult operation and the technical level needs to be improved, etc.

Pending Publication Date: 2020-07-17
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is known that MHC class I molecules are the most important molecules related to immune rejection, and knocking out such molecules is one of the breakthroughs to solve the problem of immune rejection. The direct knockout operation is extremely difficult, and the technical level needs to be improved

Method used

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  • Induced pluripotent stem cell line for reducing cell immunogenicity and construction method thereof
  • Induced pluripotent stem cell line for reducing cell immunogenicity and construction method thereof
  • Induced pluripotent stem cell line for reducing cell immunogenicity and construction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Screening and line building of the cells of Example 1

[0023] 1) Cell culture

[0024] hiPSCs culture

[0025] (1) Petri dish coating

[0026] a) Dilute Matrigel with DMEM / F12, the specific operation is as described above, and store in a refrigerator at 4°C for a short time.

[0027] b) Coat the culture dish with the prepared Matrigel one day before use, 1ml per well of the 6-well plate, seal the culture dish with parafilm and place it in a 4-degree refrigerator.

[0028] (2) Cell Recovery

[0029] a) Take out the rewarmed culture dish, suck out the Matrigel with a pipette, and quickly add 2ml of stem cell culture medium to each well;

[0030] b) Take the frozen hiPSCs out of the liquid nitrogen tank, thaw them in a 37°C water bath, and shake them gently to speed up the thawing;

[0031] c) Add 1ml of culture medium to the cryopreservation tube to neutralize the cryopreservation solution, pipette once or twice slowly, and add the cells to the culture dish at an ap...

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Abstract

The invention belongs to the field of stem cell regeneration medicine and transformation medicine, and relates to an induced pluripotent stem cell line for reducing cell immunogenicity and a method thereof. According to the invention, a human induced pluripotent stem cell line with knocking out of a beta-2 microglobulin ([beta]2m) gene (hiPSCs-[beta]<2>M<- / ->), a human induced pluripotent stem cell line with knocking out of a MHC class II transactivator (C[II]TA) gene (hiPSCs-C[II]TA<- / ->), and a human induced pluripotent stem cell line with simultaneous knocking out of the [beta]2m gene and the C[II]TA gene (hiPSCs-[beta]<2>M<- / ->&C[II]TA<- / ->) are obtained through a human cell reprogramming method. The cell line provided by the invention is a human-derived cell and has similar biologicalcharacteristics with human embryonic stem cells, and immunogenicity of human induced pluripotent stem cells can be obviously reduced after relevant immune genes are knocked out. The invention breaksaway from political and ethical disputes around use problems of human embryonic stem cells, and provides a new thought and method for treating cardiovascular diseases by stem cell transplantation.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to an induced pluripotent stem cell line that reduces cell immunogenicity and a method for establishing it, in particular to an induced pluripotent stem cell line that knocks out human leukocyte antigen (MHC)-related genes, and the induced pluripotent stem cell line Stem cell lines are cell lines with reduced immunogenicity of the cells. Background technique [0002] In recent years, the prevalence of cardiovascular-related diseases has been increasing and has become one of the most important causes of death in the world. Studies have shown that microcirculatory system lesions have a serious impact on the occurrence and development of cardiovascular diseases. Therefore, the establishment of microvascular models in vitro is of great importance. It is of great significance to study and treat cardiovascular diseases. The prior art discloses that cell programming is to reverse the di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/90C12N15/85
CPCC12N5/0696C07K14/47C12N15/907C12N15/85C12N2800/107
Inventor 陈思锋王小凯陆萌
Owner FUDAN UNIV
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