Method for inducing mononuclear cells to differentiate into dendritic cells

A technology of dendritic cells and monocytes, applied in the field of inducing the differentiation of monocytes to dendritic cells, can solve the problems of low DC induction rate, complicated operation, influence on dendritic cell differentiation, etc.

Pending Publication Date: 2021-07-30
MIAOSHUN SHANGHAI BIOTECH CO LTD
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] At present, DC cells are mainly obtained by differentiation of monocytes extracted from peripheral blood and umbilical cord blood by adding cytokines, and the induction rate of DCs obtained after differentiation is generally low, and more c

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  • Method for inducing mononuclear cells to differentiate into dendritic cells
  • Method for inducing mononuclear cells to differentiate into dendritic cells
  • Method for inducing mononuclear cells to differentiate into dendritic cells

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Embodiment

[0062] The present invention is specifically described below by way of examples. It is necessary to point out that the following examples are only used to further illustrate the present invention, and can not be interpreted as limiting the protection scope of the present invention, some non-essential improvements made by those skilled in the art according to the content of the present invention above And adjustments still belong to the protection scope of the present invention.

[0063] The embodiment provides a method, including: sorting of CD14+ monocytes, differentiation of CD14+ monocytes and flow detection;

[0064] Sorting of CD14+ monocytes:

[0065] (1) will 1×10 8 PBMC cells were mixed with 800 μL of buffer solution, and then 200 μL of CD14 magnetic beads were added to incubate in the dark at 2-8°C for 15 minutes, then 15 mL of buffer solution was added, centrifuged at 300 g for 10 minutes, the supernatant was discarded, and 500 μL of buffer solution was added to th...

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Abstract

The invention relates to the technical field of cell culture, in particular to a method for inducing mononuclear cells to differentiate into dendritic cells. The method comprises the following steps: adding CD14+ mononuclear cells into a complete culture medium I, conducting culturing for 4-5 days, adding the cultured system into a complete culture medium II, and conducting culturing for 1-2 days to obtain the dendritic cells. By controlling the sorting method of the CD14+ mononuclear cells, the number of the non-CD14+ mononuclear cells in the adhesion cells and the PBMC is effectively reduced, the content of the cells capable of being differentiated into the dendritic cells is increased, and meanwhile higher dendritic cells can be obtained under the conditions of short time and low cell factor content. GM-CSF or interleukin can be used as a cell factor to promote differentiation of dendritic cells, when GM-CSF or interleukin, such as interleukin-4, is purely added, the induction rate of DC cells is low, and when GM-CSF and interleukin-4 act together, the induction rate is high and the differentiation time is shortened under the condition of low final concentration of the cell factor.

Description

technical field [0001] The invention relates to the technical field of cell culture, and more specifically, the invention relates to a method for inducing monocytes to differentiate into dendritic cells. Background technique [0002] Dendritic cells (DC) are named for their stellate pleomorphism or dendritic processes on their surface. Dendritic cells (DC) are professional antigen-presenting cells with the strongest function in the body. cells, APC), which can efficiently uptake, process and present antigens, immature DC has a strong ability to migrate, mature DC can effectively activate initial T cells, and is at the center of initiating, regulating, and maintaining immune responses. [0003] DCs originate from bone marrow pluripotent hematopoietic stem cells, and there are two main ways of differentiation: ① Myeloid stem cells differentiate into DCs under the stimulation of GM-CSF, called myeloid dendritic cells (MDCs), also known as DC1, and single Nuclear cells and gran...

Claims

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Application Information

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IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2501/22C12N2501/23C12N2501/2304C12N2509/10
Inventor 吕丽萍俞敏娇鲍巧燕顾金海王雨婷
Owner MIAOSHUN SHANGHAI BIOTECH CO LTD
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