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Human induced pluripotent stem cell and preparation method and application thereof

A technology for pluripotent stem cells and somatic cells, which is applied in the field of human induced pluripotent stem cells and their preparation, and can solve problems such as simulation

Pending Publication Date: 2022-01-04
SHANXI MEDICAL UNIV +1
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  • Claims
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Problems solved by technology

Animal models can be used to study the pathological changes and disease mechanisms of various diseases, but the biological differences between animals and humans prevent many disease states from being well simulated by animal models

Method used

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  • Human induced pluripotent stem cell and preparation method and application thereof
  • Human induced pluripotent stem cell and preparation method and application thereof
  • Human induced pluripotent stem cell and preparation method and application thereof

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Experimental program
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Embodiment 1

[0053] In the early stage of the inventors, the type of genetic mutation in Shanxi area, which was detected in a B-type Hemophilia patient in the Second Hospital of Shanxi Medical University, and the F9 gene mutation C.223C> T was confirmed (HTTP) : / / www.factorix.org / This mutation is a variable mutation.

[0054] First, people induce multi-capable cell preparation

[0055] Collect urine cells:

[0056] With the patient's consent, the urine peeled cells were separated from the urine sample of hemophilia, proliferated by urine peeled cells [Zhou T, Benda C, Dunzinger S, et al.generation ofhuman inducedPluripotent Stem Cells from Urine Samples " ] .Nature Protocols, 2012,7 (12): 2080-2089.

[0057] The middle stage was collected, then 400 g at room temperature was centrifuged for 10 minutes to separate human urine cells (HUCs). Cell deposition was washed twice with DPBS (GIBCO). The primary urine cell culture was cultured in urine medium, which was mixed by a DMEM base medium 1: 1 ...

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Abstract

The invention relates to a human induced pluripotent stem cell strain SXMUi002-A-1. The human induced pluripotent stem cell strain SXMUi002-A-1 is preserved in the China Center for Type Culture Collection, and the preservation number of the human induced pluripotent stem cell strain SXMUi002-A-1 is CGMCC No: 22359. The iPSC is obtained by extracting urine cells from urine of a B-type hemophilia patient carrying F9c.223C>T(p.R75X) nonsense mutation and reprogramming, and further research finds that the iPSC is similar to an embryonic stem cell in the aspects of cellular morphology, gene expression, proliferation and differentiation potential and has the potential of differentiating to three germ layers. The prepared human induced pluripotent stem cell strain SXMUi002-A-1 has important value for molecular mechanism research of hemophilia nonsense mutation, hemophilia detection, drug screening, gene therapy and other clinical application researches, and provides important resources.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a human induced pluripotent stem cell and a preparation method thereof. Background technique [0002] Hemophilia is a X-chromosome-chain hidden hereditary hereditary disease. Common characteristics is that activated coagulinary enzyme generation disorders, coagulation time is prolonged, and life is tenden to have a slight trauma, and there is no obvious trauma in severe cases. Sex "bleeding. F8 gene defects lead to hemophilia A (Hemophilia), F9 gene defects, lead to hemophilia B (HB), in the male population, the incidence rate is 1 / 5,000 and 1 / 50,000 [Yang Renchi. Hemophilia Diagnosis and Treatment of Chinese Expert Consensus (2017 Edition) [J]. Chinese Journal of Hematology, 2017,38 (05): 364-370. Factorix Gene (f9) Variant Database (http: / / www.factorix.org / ) shows 1094 F9 variants in 3713 cases, of which 87 are sanctuous, with 8%. [0003] The existing treatment means of hemop...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N5/071C12N15/85
CPCC12N5/0696C12N15/85C12N2501/603C12N2501/602C12N2501/604C12N2501/606C12N2506/25
Inventor 王刚杨林花马艳春姚明泽吴长新刘秀娥任娟张睿娟张夏林孙雯雯赵丽东
Owner SHANXI MEDICAL UNIV
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