Co-culture method for in-vitro activation of DC cells through heat shock protein gp96-polypeptide complex

A technology of heat shock protein and polypeptide complex, which is applied in the field of co-cultivation of heat shock protein gp96-polypeptide complex to activate DC cells in vitro, which can solve the problems of ineffective effect and low efficiency, so as to improve activation efficiency and enhance cell activity , the effect of enhancing the ability to absorb antigens

Pending Publication Date: 2021-05-14
和泓尚医(成都)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, in the existing technology, the use of biotechnology to activate DC cells mainly through in vivo biological pathways, the efficiency is not high, and the effect is not obvious

Method used

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  • Co-culture method for in-vitro activation of DC cells through heat shock protein gp96-polypeptide complex
  • Co-culture method for in-vitro activation of DC cells through heat shock protein gp96-polypeptide complex
  • Co-culture method for in-vitro activation of DC cells through heat shock protein gp96-polypeptide complex

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Embodiment

[0036] Example: as attached Figure 1-9 As shown, the present invention is a co-cultivation method for heat shock protein gp96-polypeptide complex to activate DC cells in vitro, comprising steps:

[0037] S1. Prepare 50 mL of DC cell induction medium (containing 45 mL of DC medium, 5 mL of DC medium supplement, 15 μL of DC-I and 5 μL of DC-II);

[0038] Notice:

[0039] ①. For DC medium additives, put them in the -20°C refrigerator in advance and thaw slowly;

[0040] ② After the DC medium additive is completely dissolved, shake the bottle gently to make the temperature and composition uniform. When shaking, be careful to avoid air bubbles; when thawing, if there is protein precipitation, remove it by centrifugation or use a 0.22um filter.

[0041] S2. On day 0 of culture, prepare 1*10 8 PBMC cells were resuspended with 50 mL of RPMI1640 culture medium, spread in a T75 culture flask (that is, the culture flask was covered with 50 mL of PBMC suspension), and incubated in a C...

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Abstract

The invention discloses a co-culture method for in-vitro activation of DC cells through a heat shock protein gp96-polypeptide complex, and mainly relates to the field of biological medicine. The method comprises the following steps: S1, preparing a DC cell induction culture medium; S2, in the 0 day of culturing, resuspending by using an RPMI1640 culture solution, standing in a CO2 incubator, and incubating for 2 hours; S3, slightly shaking a culture flask, removing the suspended cell sap, washing, and continuing to culture in the CO2 incubator; S4, in the second day of culturing, adding DC-III, DC-IV and DC-V into the culture flask; S5, in the third day of culturing, collecting the DC cells in a centrifugal tube, and carrying out centrifugal treatment and resuspension; S6, adding the heat shock protein gp96-polypeptide complex reagent into an EP tube in the step S5. In-vitro activation efficiency of the DC cells can be improved by the method, the antigen uptake capacity of the DC cells is enhanced, and the activation efficiency of specific T cells is improved.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a co-cultivation method for the heat shock protein gp96-polypeptide complex to activate DC cells in vitro. Background technique [0002] Dendritic cells (dendritic cells, DCs) were discovered by Steinman and Cohn in 1973 and are named for their stellate pleomorphism or dendritic protrusions on their surface. DC has morphological characteristics and motility that other cells do not have, and is widely distributed. DC not only has phagocytosis ability, but also is the most powerful antigen-presenting cell known so far, which can effectively stimulate the activation of T lymphocytes and B lymphocytes , so as to organically link innate immunity and acquired immunity, and play an important role in the treatment of diseases such as tumors. [0003] Placenta-derived polypeptides contain a large number of heat shock proteins. Heat shock protein (HSP) is a type of protein that is highly conser...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/998
Inventor 王刚范志青郑清富
Owner 和泓尚医(成都)生物科技有限公司
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