Preparation method of pluripotent stem cells of xeroderma pigmentosum patient

A technology of pluripotent stem cells and coloration, which is applied in the field of preparation of pluripotent stem cells and nerve cells, can solve problems such as difficulty in obtaining experimental materials, and achieve great application prospects

Active Publication Date: 2016-05-11
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the mechanism of neurodegeneration in patients with pigmented dry skin is stagnant, the main reason is that it is difficult to obtain suitable experimental materials

Method used

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  • Preparation method of pluripotent stem cells of xeroderma pigmentosum patient
  • Preparation method of pluripotent stem cells of xeroderma pigmentosum patient

Examples

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Effect test

Embodiment 1

[0104] Example 1. A method for preparing neural stem cells and neurons with specific gene mutations for patients with pigmented dry skin

[0105] 1. Acquisition and identification of induced pluripotent stem cells

[0106] 1. Acquisition of induced pluripotent stem cells

[0107] The fibroblasts derived from pigmented dry skin patients carrying different gene mutations were reprogrammed to obtain induced pluripotent stem cells with XPA mutations, induced pluripotent stem cells with XPB mutations, induced pluripotent stem cells with XPC mutations, and induced pluripotent stem cells with XPG mutations. Induced pluripotent stem cells and XPV-mutated induced pluripotent stem cells. The specific method is as follows:

[0108] (1) Cell recovery and expanded culture

[0109] The skin fibroblasts derived from the following five strains of pigmented dry skin patients carrying different gene mutations: XPA fibroblasts (GM00710), XPB fibroblasts (GM13026), XPC fibroblasts (GM15709), X...

Embodiment 2

[0153] Example 2. Verification of UV Sensitivity of Neural Stem Cells and Neurons Carrying Patient-Specific Gene Mutations of Pigmented Xeroderma

[0154]After UV irradiation, DNA mainly exists in two forms of damage, which are cyclobutanepyrimidine dimers (CPD) and 6-4 photoproducts (6-4pyrimidine photoproducts, 6-4PPs). These two forms are completely dependent on the intracellular nucleotide splicing repair pathway for repair, so the content of CPD can be used to indicate the ability of cells to repair nucleotide splicing. In this example, a certain dose of UV was used to irradiate the five kinds of neural stem cells and neurons carrying the patient-specific gene mutation of pigmented dry skin prepared in Example 1, and the content of CPD in the cells and the biological response of the cells were detected.

[0155] 1. UV Sensitivity Verification of Neural Stem Cells Carrying Specific Gene Mutations of Patients with Pigmented Dry Skin

[0156] 1. Detection of CPD content and...

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Abstract

The invention discloses a preparation method of pluripotent stem cells of a xeroderma pigmentosum patient. The method comprises the steps that by means of an iPSC technology, skin fibroblast cells of the xeroderma pigmentosum patient are reprogrammed into specific induced pluripotent stem cells through a non-integration episomal plasmid electrophoretic transfer method and directionally divided into nerve cells which show extreme sensitivities to UV in vitro, cell apoptosis is prone to occurrence, and some clues are provided for neurodegenerative diseases of the xeroderma pigmentosum patient. In addition, the mutational neural stem cells and neurones which carry genes of the patient can serve as an effective platform to conduct efficient and high-throughout personalized drug screening, and a foundation is laid for disease study, disease model development, research on pathogenesis of diseases and treatment on the diseases. A huge application prospect is achieved in personalized treatment and translational medicine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing pluripotent stem cells from patients with pigmented dry skin, in particular to a method for preparing various specific pluripotent stem cells and nerve cells of patients with pigmented dry skin. Background technique [0002] Xeroderma pigmentosum (XP for short) is a rare autosomal recessive genetic disease caused by gene mutations leading to DNA repair defects. According to the difference of the corresponding mutant genes (XPA, XPB, XPC, XPD, XPE, XPF, XPG and XPV), patients with xeroderma pigmentosa can be divided into the following 8 different complementary types: xeroderma pigmentosa complementary type A, Xeroderma Pigmento Complementary B, Xeroderma Pigmento Complementary C, Xeroderma Pigmento Complementary D, Xeroderma Pigmento Complementary E, Xeroderma Pigmento Complementary F, Xeroderma Pigmento Complementary type G and xeroderma pigmentosa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12Q1/02
CPCC12N5/0619C12N5/0623C12N2501/602C12N2501/603C12N2501/604C12N2501/606C12N2501/608C12N2506/1307C12N2506/45G01N33/5058G01N33/5073G01N2500/10
Inventor 刘光慧曲静付丽娜任若通
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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