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Method for establishing endothelial crossing model for asthmatic serum dendritic cell

A technology of dendritic cells and serum, applied in the field of medical models, can solve problems such as lack of models

Inactive Publication Date: 2010-01-27
周林福
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Problems solved by technology

Therefore, there is a lack of an experimental platform model, and experiments that truly simulate the differentiation, maturation and migration of monocytes in patients with asthma cannot be performed

Method used

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  • Method for establishing endothelial crossing model for asthmatic serum dendritic cell
  • Method for establishing endothelial crossing model for asthmatic serum dendritic cell

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Embodiment Construction

[0021] The method for establishing the endothelial crossing model of serum dendritic cells in asthma is as follows:

[0022] (1) mononuclear cells were obtained from human peripheral blood by immunomagnetic bead method;

[0023] (2) Obtain fresh umbilical cords from normal puerperas (within 4 hours from the delivery room), trypsinization method (0.25% trypsin) to obtain human umbilical vein endothelial cells from the umbilical cord veins of normal puerperas, the optimal digestion time is 20 minutes, primary Subculture when the cultured cells are 70% confluent;

[0024] (3) After the cultured endothelial cells grew into a monolayer, the endothelial cells showed a paving stone-like appearance under a phase contrast microscope, and the cytoplasm of the endothelial cells stained positively for human factor VIII under a fluorescence microscope;

[0025] (4) 24-well cell culture plate was pretreated with 0.1% gelatin, and the passaged (second generation) endothelial cell suspension...

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Abstract

The invention relates to a method for establishing an endothelial crossing model for asthmatic serum dendritic cells. The method comprises the following steps: adopting Ia centrifugation method to obtain serums, an immunomagnetic bead separation to obtain monocytes, and a trypsin digestion method to obtain human umbilical venous endothelial cells; pretreating the serums, the monocytes and the human umbilical venous endothelial cells with gelatin, adding passage endothelial cell suspension into a culture plate to grow into a single layer, suspending asthmatic patient serums on the monocytes to be added on the endothelial cell single layer, fully washing the endothelial cell single layer, and recovering suspension cells to obtain the endothelial crossing model for the asthmatic serum dendritic cells. The invention overcomes the defect that an experiment for truly simulating differentiation maturation and migration functions of the monocytes in the body of an asthmatic patient can not be carried out because of one experiment platform short. The invention provides an in vitro model depended by non cytokines for objectively simulating the monocytes to migrate and differentiate in the body to become the dendritic cells, and in addition, the asthmatic serums are a true internal environment of the patient.

Description

technical field [0001] The invention relates to a medical model, in particular to a method for establishing an asthmatic serum dendritic cell endothelial crossing model. Background technique [0002] Asthma is CD4 + In airway allergic diseases mediated by T cells, eosinophil (EOS) infiltration is a prominent pathological feature, and type II helper T cell (Th2) immune hyperactivity is the most important immunological abnormality. Dendritic cells (dendritic cells, DCs) are the initiators and maintainers of immune responses, and play a key role in the process of Th2 immune drift in asthma. [0003] Monocytes are the common precursor cells of macrophages and DCs, and can differentiate into DCs under the induction of various exogenous cytokines. In the classic granulocyte-macrophage colony-stimulating factor (GM-CSF) / interleukin-4 (IL-4) system, it takes about 5 days for monocytes to differentiate into immature DCs, followed by the maturation inducer lipopolysaccharide ( LPS)...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12Q1/02
Inventor 周林福殷凯生顾晓燕张明顺季晓辉
Owner 周林福
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