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Method for preparing plasmocyte and plasmocytes obtained by method

A technology for plasma cells and B cells, applied in the field of preparation of plasma cells, can solve the problems of not being able to simultaneously improve the ability of plasma cells to secrete antibodies, the ability of plasma cells to secrete antibodies in general, and many cells to die, and achieve short induction time and strong ability to secrete antibodies. , the effect of high purity

Inactive Publication Date: 2012-12-05
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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Problems solved by technology

[0004] At present, B cells are mostly isolated from the spleen of mice and stimulated with LPS in vitro to obtain plasma cells, which have the following limitations: it takes a long time, more cells die during the induction process, and the ability of the generated plasma cells to secrete antibodies is average
Although it can promote plasma cell differentiation by interfering with the expression of transcription factors that inhibit plasma cell differentiation, it cannot simultaneously improve the antibody secretion capacity of plasma cells

Method used

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  • Method for preparing plasmocyte and plasmocytes obtained by method
  • Method for preparing plasmocyte and plasmocytes obtained by method
  • Method for preparing plasmocyte and plasmocytes obtained by method

Examples

Experimental program
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Embodiment 1

[0042] Example 1, preparation of plasma cells and performance verification of plasma cells

[0043] 1. Take the mouse spleen, cut it into pieces, grind it, and use Ficoll density gradient centrifugation (the density of Ficoll used is 1.073g / ml) to separate mononuclear cells, and then use the Mini MACS B220 cell immunomagnetic bead sorting system (Germany Miltenyi Company, Cat.No.130-049-501) to isolate B cells (B220+B cells) positive for the cell surface marker B220.

[0044] 2. Suspend the B220+B cells obtained in step 1 in RPMI1640 medium to obtain 2×10 5 cells / ml of cell suspension.

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Abstract

The invention discloses a method for preparing a plasmocyte and the plasmocyte obtained by the method. The invention provides a method for preparing the plasmocyte from a B cell. The method comprises the following steps of: (1) interfering the expression of an MYSM1 gene in the isolated B cell, wherein the MYSM1 gene is a gene of protein which is shown in a sequence 1 of a coding sequence table; and (2) inducing the cell by using lipopolysaccharide, so that the cell is differentiated to form the plasmocyte. The method has the advantages that (1) the plasmocyte is high in purity, multiple in number and high in capacity of secreting antibodies; and (2) the method is short in induction time, easy to operate, convenient and practical. The stable method for preparing the plasmocyte for secreting the antibodies in high efficiency is established, so that the foundation is established for the study and application of the plasmocyte and the antibodies thereof.

Description

technical field [0001] The invention relates to a method for preparing plasma cells and the obtained plasma cells. Background technique [0002] After receiving antigen stimulation, mature B cells become activated B cells with the assistance of antigen-presenting cells and helper T cells, and then differentiate into plasma cells, which synthesize and secrete various immunoglobulins. [0003] Plasma cells and the antibodies they secrete play an important role in the development of protective immunity, tumors and autoimmune diseases. [0004] At present, B cells are mostly isolated from mouse spleen and stimulated with LPS to obtain plasma cells in vitro, which has the following limitations: it takes a long time, more cells die during the induction process, and the ability of the generated plasma cells to secrete antibodies is average. Although plasma cell differentiation can be promoted by interfering with the expression of transcription factors that inhibit plasma cell diff...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10
Inventor 江小霞毛宁
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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