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DC cell culture method, culture medium, DC treatment strategy-based drug and application of tyrosine kinase inhibitor in preparation of DC cell culture medium

A tyrosine kinase and cell culture technology, which is applied in the field of preparation of tyrosine kinase inhibitors, can solve the problems that DC cell culture methods cannot meet, and achieve high maturity or high activity, increase maturity or Activity, effect of improving expression

Pending Publication Date: 2021-05-28
SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention discloses a DC cell culture method, a culture medium, a drug based on a DC treatment strategy, and the application of a tyrosine kinase inhibitor in the preparation thereof, so as to solve the problem that the current DC cell culture method cannot meet the requirements of high maturity or high degree of clinical efficacy. The problem of active DC cells, to achieve the purpose of improving the maturity or activity of DC cells

Method used

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  • DC cell culture method, culture medium, DC treatment strategy-based drug and application of tyrosine kinase inhibitor in preparation of DC cell culture medium
  • DC cell culture method, culture medium, DC treatment strategy-based drug and application of tyrosine kinase inhibitor in preparation of DC cell culture medium
  • DC cell culture method, culture medium, DC treatment strategy-based drug and application of tyrosine kinase inhibitor in preparation of DC cell culture medium

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Embodiment 1

[0031] An embodiment of the present invention provides a DC cell culture method, which includes the following specific steps:

[0032] First, the peripheral blood mononuclear cells were adhered to the wall for 2 hours, and then the adherent cells were left to be cultured in DCI medium, which was supplemented with 5% human serum and 800 U / mL of GM-CSF (grains cells-macrophage colony-stimulating factor) and 500 U / mL IL-4 (interleukin-4) in the basal medium, after induction and culture for 7 days, imDC cells (immature DC cells) were obtained.

[0033] Then, 10 ng / mL of TNF-α (tumor necrosis factor-α, tumor necrosis factor-α), 1 mg / mL of PGE2 (prostaglandin E2) and 1000 U / mL of IL-6 were added to the above DCI medium. (interleukin-6) and 25nM dasatinib, and continued to culture for 3 days to obtain mDC cells (mature DC cells).

[0034] Among them, GM-CSF can promote the development of myeloid cells and is the most fundamental cytokine for maintaining and inducing DC development a...

Embodiment 2

[0041] Embodiment of the present invention has carried out comparative test, specifically as follows:

[0042] 1. Obtain human peripheral blood mononuclear cells

[0043] 20ml of anticoagulated peripheral blood was extracted from healthy donors (HD, n=10) and chronic myelogenous leukemia patients with MR4.5 remission (CML, n=10) who received imatinib treatment, and respectively mixed with PBS (Phosphate-buffered saline) was diluted 1:1, and then slowly added 400g of lymphocyte separation medium accordingly, and centrifuged at 4°C for 30 minutes to collect the white cell layer; then, wash the buffy coat with PBS to obtain PBMC (peripheral blood mononuclear cells).

[0044] 2. Generation and maturation of DC cells

[0045] Each example of PBMC obtained above was divided into 2×10 6 The concentration of cells per ml was determined by RPMI 1640 (RPMI is the abbreviation of Roswell Park Memorial Institute, referring to Roswell Park Memorial Institute; RPMI 1640 refers to a cell ...

Embodiment 3

[0081] The embodiment of the present invention provides a drug based on a DC treatment strategy, such as a DC vaccine and a drug for DC cell therapy, wherein the Dasatinib component can be added as an adjuvant for enhancing the DC vaccine and for DC cells The efficacy of therapeutic drugs.

[0082] Of course, other tyrosine kinase inhibitors such as imatinib or nilotinib can also be used in the above drugs based on the DC treatment strategy instead of dasatinib as an adjuvant component for enhancing drug efficacy.

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Abstract

The invention discloses a DC cell culture method, a culture medium, a DC treatment strategy-based drug and application of a tyrosine kinase inhibitor in preparation of the DC cell culture medium. According to the DC cell culture method, in the culture process of induced differentiation of immature DC cells, a tyrosine kinase inhibitor is added into a culture medium, and the mature DC cells are obtained through induced differentiation culture; the culture medium is a cell culture medium containing a tyrosine kinase inhibitor and can be used for culturing immature DC cells which are induced to be differentiated into mature DC cells; the medicine based on the DC treatment strategy contains a tyrosine kinase inhibitor; and the tyrosine kinase inhibitor can be used in the preparation of the DC treatment strategy-based medicine. According to the scheme, the problem that an existing DC cell culture method cannot meet the requirement for highly mature or high-activity DC cells required by clinical curative effects can be solved, and the aim of improving the maturity or activity of the DC cells is achieved.

Description

technical field [0001] The invention relates to the related technical field of DC cells, in particular to a method for culturing DC cells, a culture medium, a drug based on a DC treatment strategy, and an application of a tyrosine kinase inhibitor in the preparation thereof. Background technique [0002] DC cells (dendritic cells), as the most important antigen-presenting cells, can play an important role in anti-tumor immunity by priming naive T cells and promoting their proliferation and differentiation into Th cells and cytotoxic T lymphocytes (CTL) . [0003] In the past few decades, anti-tumor treatment strategies based on DC cells have developed rapidly; for example, cultivating autologous dendritic cells to enhance their activity, and then reinfusing them into patients is the cutting-edge technology of current research and application. Not only can it effectively stimulate the anti-tumor immune response in tumor patients, and it has shown good curative effect in rese...

Claims

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Application Information

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IPC IPC(8): C12N5/0784A61K39/00A61K35/17A61P35/02
CPCC12N5/0639A61K39/001191A61K35/17A61P35/02C12N2501/22C12N2501/2304C12N2501/727C12N2501/25C12N2501/2306C12N2501/02C12N2501/999
Inventor 代景莹何林夏静仪曹婉君
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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