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Novel efficient method for separating human T-Lymphocytes by immunomagnetic beads

An immunomagnetic bead sorting and lymphocyte technology, which is applied in the field of cell culture, can solve the problem of the decrease in the number of viable cells, achieve the effect of prolonging the survival time and being simple and easy to operate

Inactive Publication Date: 2012-10-10
赵品楠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The culture of T lymphocytes belongs to the primary culture. Due to the inability to be passaged, the number of living cells used in the experiment gradually decreases with the prolongation of the experiment time.

Method used

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  • Novel efficient method for separating human T-Lymphocytes by immunomagnetic beads

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Experimental program
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Embodiment Construction

[0008] 1. Materials and methods

[0009] 1.1 Research object

[0010] T lymphocytes of healthy adults (mainly screening healthy, disease-free volunteer blood donors).

[0011] 1.2 Materials

[0012] Positive selection human CD3 kit (StemCell Company), human lymphocyte separation medium (Tianjin Haoyang Biological Products Technology Co., Ltd.), RPMI-1640 cell culture medium (Chengdu Harry Biotechnology).

[0013] 1.3 Isolation and culture of human peripheral blood mononuclear cells

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Abstract

The invention establishes an operation method in T-Lymphocytes culture in vitro by immunomagnetic bead separation technique to maximize the separation of human T-Lymphocytes from the same volume of human peripheral blood to obtain a lot of purified T-Lymphocytes and cover the shortage that lymphocytes can be only primary cultured but not be subcultured to the greatest extent. The method is simple and practicable.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to the sorting of human T lymphocytes and the operation method of the immunomagnetic bead separation method. Background technique [0002] Immunomagnetic bead sorting is an advanced cell separation technology widely used in scientific research. It combines cell surface antigen molecules with specific monoclonal antibodies connected to magnetic beads, and under the action of an external magnetic field, cells connected to magnetic beads through monoclonal antibodies are attracted and retained in the magnetic field; without this surface antigen Cells, because they cannot bind to the specific monoclonal antibody attached to the magnetic beads, cannot stay in the magnetic field, thereby separating the cells. As cells grown in suspension, T lymphocytes cannot be isolated from adherent cells by adherent selection. Therefore, the method of sorting T lymphocytes can be mostly used i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 赵品楠
Owner 赵品楠
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