Method for mutagenizing mice lymphocyte

A technology of lymphocytes and cells, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of limiting the clinical application of iPS cells, and achieve the effects of avoiding potential harm, wide sources, and materials

Inactive Publication Date: 2012-10-03
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, the study found that the generation of iPS cells requires virus-mediated defined genes to be successful
Therefore, viruses as potential carcinogens limit the clinical application of iPS cells

Method used

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  • Method for mutagenizing mice lymphocyte

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Embodiment Construction

[0019] 1 Test materials and methods

[0020] 1.1 Isolation of lymphocytes

[0021] Take green fluorescent protein transgenic mice, and the green fluorescent protein transgenic mice are commercialized transgenic mice. They are killed by neck dislocation, and the abdomen is disinfected with alcohol. Gently grind the cells in the mesh nylon omentum to form single cells, and collect after washing with PBS, centrifuge to remove the supernatant, and resuspend in serum-free DMEM culture medium for later use.

[0022] 1.2 Culture of mouse ES cells

[0023] Fetal fibroblasts from 12.5-day-old mice were cultured with high-sugar DMEM medium. After the cells covered the culture dish, they were digested and subcultured with cell digestion solution containing 2.5 mg / ml trypsin-0.2 mg / ml EDTA. The fetal fibroblasts in the culture dish were treated with 10 μg / ml mitomycin for 2.5 hours, digested with the cell digestion solution containing 2.5 mg / ml trypsin-0.2 mg / ml EDTA, and seeded into th...

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Abstract

The invention discloses a method for mutagenizing a mice lymphocyte. The method comprises the following steps: firstly, separating lymphocyte; secondly, culturing a mice ES cell; thirdly, fusing the lymphocyte with the ES cell; and fourthly, carrying out separation culture on a mutagenic cell. The lymphocyte is used as an important component cell in an animal body and has the advantages of wide sources, easily-obtained raw materials and the like. According to the method disclosed by the invention, the mice lymphocyte is induced to form a stem cell by means of a cell fusion technology; the formed stem cell has the characteristics of the morphology, gene expression, the in vitro differentiation capacity and the like endowed by the mice ES cell, and thus potential hazards of viruses in stem cell induction are avoided. Therefore, the method lays a foundation for wide application of transforming the mice somatic cell into the stem cell and also provides important reference for research and utilization of stem cells of animals or human beings.

Description

[0001] technical field [0002] The invention relates to a mouse lymphocyte mutagenesis method. Background technique [0003] Stem cells, as the origin cells of the body composition, are primitive undifferentiated cells with multi-directional differentiation potential and self-replication ability, and are the ancestor cells that form various tissues and organs of mammals. Stem cells usually have common features in shape, presenting a round or oval shape, stem cells are small in size, and their nuclei are relatively tall. Stem cells, as a kind of cells that have not yet fully differentiated and matured, have the potential to form cells of various tissues and organs under induced environmental conditions, and are called "universal cells" by the medical community. Stem cells have a wide range of uses and involve many fields of medicine. Stem cells have strong self-renewal and proliferation capabilities, and have become the carrier cells of transgenes. Embryonic stem cells (E...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/06C12N5/071
Inventor 曹鸿国章孝荣杨盼蒲勇张运海刘亚陶勇方富贵李运生
Owner ANHUI AGRICULTURAL UNIVERSITY
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