A special medium and culture method for inducing mouse bone marrow cells to stably differentiate into immature dendritic cells

A technology of dendritic cells and bone marrow cells, applied in the special medium and culture field of immature dendritic cells, can solve the problem of unstable imDCs quantity and maturity, unable to obtain sufficient imDCs, affecting accuracy and reliability, etc. problem, to achieve the effect of relatively simple fixation of ingredients, optimization of identity, and cost savings

Active Publication Date: 2021-06-18
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the limitation of clinical blood samples, the low self-proliferation ability of DCs, and the large differences and complex components between different brands of media and serum reagents, it is impossible to obtain sufficient imDCs, the culture efficiency is low, the cell activity is poor, and each time The amount and maturity of imDCs obtained are unstable, which seriously affects the accuracy and reliability of subsequent experiments

Method used

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  • A special medium and culture method for inducing mouse bone marrow cells to stably differentiate into immature dendritic cells
  • A special medium and culture method for inducing mouse bone marrow cells to stably differentiate into immature dendritic cells
  • A special medium and culture method for inducing mouse bone marrow cells to stably differentiate into immature dendritic cells

Examples

Experimental program
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Effect test

Embodiment 1

[0044] The DC culture medium of this example includes the following components: X-VIVO 20 95v / v%, LPS 1 μg / ml, GM-CSF 40ng / ml, IL-4 10ng / ml, penicillin-streptomycin 10 6 U / ml, the balance is sterile water for injection.

[0045] The method for culturing and inducing mouse bone marrow cells to differentiate into imDCs using the DC medium described in this example comprises the following steps:

[0046] S1. Bone marrow cells collected from the tibia and femur of 6-8-week-old mice were collected by RPMI1640 in an ice bath environment and made into a suspension, centrifuged at 1000rpm at 4°C for 5 minutes, and discarded the supernatant;

[0047] S2. Add 3-5ml of erythrocyte lysate to the cell pellet obtained by centrifugation in step S1, mix well and let stand for 2 minutes, centrifuge at 1000rpm at 4°C for 2 minutes and discard the supernatant;

[0048] S3. Resuspend the cells centrifuged in step S2 in RPMI1640, centrifuge at 1000 rpm for 2 minutes at 4°C and discard the superna...

Embodiment 2

[0058] The DC culture medium of this example includes the following components: X-VIVO 20 95v / v%, LPS 1 μg / ml, GM-CSF 10ng / ml, IL-4 40ng / ml, penicillin-streptomycin 10 6 U / ml, the balance is sterile water for injection.

[0059] The method for culturing DC cells using the DC medium of this example is similar to the method in Example 1.

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Abstract

The invention belongs to the technical field of cell culture, and discloses a culture medium and a culture method for inducing mouse bone marrow cells to stably differentiate into immature dendritic cells. The medium includes serum-free medium, lipopolysaccharide, granulocyte-macrophage colony-stimulating factor, interleukin-4, penicillin-streptomycin mixed in proportion; the DC culture medium of the present invention can be stably Inducing a high proportion of mouse bone marrow cells to differentiate into immature dendritic cells, the cells are in good growth state, the number, purity and biological function of the cells are stable, which can meet the needs of further experiments, and the experimental results are stable, and the cell culture medium of the present invention The production cost is low, the use is convenient, and the utility model has practical application value.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a special culture medium and culture method for immature dendritic cells. Background technique [0002] Dendritic cells are a type of antigen-presenting cells (Antigen presenting cells, APCs) that exist in peripheral blood, skin, lymphoid organs, and thymus. Therefore named dendritic cells (dendritic cells, DCs). It efficiently uptakes, processes and presents antigens and stimulates the activation of naive T cells after maturation. Immature DC cells have a strong ability to migrate, absorb and process antigens, while mature DC cells have a strong ability to stimulate the proliferation and differentiation of naive T cells, and are at the center of initiating, regulating, and maintaining immune responses. The in-depth study of the imDC maturation process will help to further clarify the mechanism of immune tolerance, which is of great significance for clinical and basic research field...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/052C12N2501/22C12N2501/2304C12N2506/11
Inventor 马毅薛志诚章旭之邓荣海吴旭坤饶佳伟周晓转
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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