Human dendritic cell induction method and composition for resisting viruses and tumors
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A technology of dendritic cells and compositions, applied in the field of immune cell anti-virus, anti-tumor, and cell culture, can solve the problems of short culture time, long cycle, weak ability to induce immune response, etc., and achieve the effect of short culture time
Active Publication Date: 2021-09-28
GUANGXI UNIV OF CHINESE MEDICINE
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[0008] The object of the present invention is to provide a method and composition for inducing human dendritic cells for anti-virus and anti-tumor. The method for inducing human dendritic cells for anti-virus and anti-tumor based on the composition induces and cultivates DCs with great potential. Strong functional, in
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Embodiment 1
[0044] Example 1 Induction and culture of mature DC
[0045] (1) Use blood collection supplies and heparin sodium anticoagulant tubes to obtain 10ml of peripheral blood from volunteers over the age of 18 without disease.
[0046] (2) Prepare biological safety cabinets and related aseptic operating supplies.
[0047] (3) Take an equal volume of 1×PBS, and mix the blood and PBS thoroughly in a centrifuge tube.
[0048] (4) Add 4ml Lymphoprep to two 15ml centrifuge tubes respectively TM For lymphocyte separation solution, slowly pour 10ml of blood and PBS mixture into a 15ml centrifuge tube along the tube wall, a total of two tubes. The lymphocyte separation solution and the mixed solution should be layered. Centrifugation: 1500rpm, 20min, centrifugal adjustment speed up 4, speed down 4.
[0050] (6) Add 9ml of buffer solution to two new 15ml centrifuge tubes, recover human ...
Embodiment 2
[0061] Example 2 Comparison with traditional mature DC culture methods
[0062] 1. Traditional method for mature DC culture
[0063] (1) Use blood collection supplies and heparin sodium anticoagulant tubes to obtain 10ml of peripheral blood from volunteers over the age of 18 without disease.
[0064] (2) Prepare biological safety cabinets and related aseptic operating supplies.
[0065] (3) Take an equal volume of 1×PBS, and mix the blood and PBS thoroughly in a centrifuge tube.
[0066] (4) Add 4ml Lymphoprep to two 15ml centrifuge tubes respectively TM For lymphocyte separation solution, slowly pour 10ml of blood and PBS mixture into a 15ml centrifuge tube along the tube wall, a total of two tubes. The lymphocyte separation solution and the mixed solution should be layered. Centrifugation: 1500rpm, 20min, centrifugal adjustment speed up 4, speed down 4.
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Abstract
The invention relates to the field of cell culture and virus and tumor resistance of immune cells, and particularly discloses a human dendritic cell induction method and composition for virus and tumor resistance. The human dendritic cell induction composition comprises a mature human dendritic cell induction composition and an immature human dendritic cell induction composition, wherein the mature human dendritic cell induction composition consists of poly I: C and recombinant human interleukin 1 beta (IL-1beta), and the immature human dendritic cell induction composition consists of a granulocyte-macrophage colony stimulating factor (GM-CSF), recombinant human interleukin 4 (IL-4) and interferon beta (IFN-beta). The human dendritic cell induction method based on the human dendritic cell induction composition is used for inducing and culturing for 3 days to obtain a DC which is short in culture period, strong in aggregation, strong in function of inducing allogenic initial CD4 + T cells and CD8 + T cells and capable of inducing antigen-specific CD8 + T cells; the culture period can be greatly shortened by utilizing the method, and the highly-functional DC is obtained and is used as an early-stage basis of virus resistance and tumor resistance of clinical cell vaccines.
Description
technical field [0001] The invention belongs to the fields of cell culture and immune cell anti-virus and anti-tumor, and particularly relates to a method and composition for inducing human dendritic cells for anti-virus and anti-tumor. Background technique [0002] 1 The role of human DC in the adaptive immune response [0003] Dendritic cells (DC) are the most powerful professional antigen-presenting cells in the human body, connecting innate immunity and adaptive immunity. DCs initiate adaptive immune responses by presenting pathogen-derived antigens to antigen-specific T cells and play a central role in adaptive immune responses and self-tolerance. DC is a heterogeneous population, divided into conventional DC (cDC), plasmacytoid DC (pDC), monocyte-derived DC (moDC), and Langerhans cells (LNs). Under normal circumstances, DC is in an immature and non-functional state, patrols in peripheral tissues, uses innate immune receptors (including Toll-like receptors) to recogni...
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