DC (dendritic cell) preparation method

A cell and granulocyte technology, applied in the field of DC cell preparation, can solve the problems of uncontrollable time and scale of DC preparation, and achieve the effects of convenient preparation method, high vitality and purity, and controllable quality

Inactive Publication Date: 2016-07-06
SHANGHAI ANGECON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Conventional DC cell preparation is to isolate PBMC cells from fresh whole blood, and then directly enter the link of DC cell preparation. The acquisition of raw whole blood is accidental and limited, and the time and scale of DC preparation are not controllable.

Method used

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  • DC (dendritic cell) preparation method
  • DC (dendritic cell) preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Recovery of PBMC cells

[0038] Preheat the water bath to 37°C, then take out the frozen PBMC cells from the liquid nitrogen at -196°C, and shake them quickly in the water bath at 37°C to make them thaw quickly.

[0039] 2. Isolation of DC cells

[0040] Gently remove the cells, adjust the mononuclear lymphocyte concentration to 2×10 with GT-T551 serum-free medium 6 cells / mL, then add the adjusted concentration of mononuclear lymphocytes into the six-well plate at 3 mL / well, adhere to the wall for 1.5 hours, and finally discard the upper layer of unattached cells in each well of the six-well plate. The lower layer of the well is left with immature DC cells.

[0041] 3. Cell inoculation

[0042] Add 3 mL of DC medium to each well of the six-well plate, place it in a carbon dioxide incubator with a carbon dioxide concentration of 5%, and a temperature of 37%. After culturing for 48 hours, add 0.75 mL of mixing factor to induce DC cells .

[0043] 4. Induced matura...

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Abstract

The present invention relates to the field of cell preparation, in particular to a method for preparing DC cells. The frozen PBMC cells are taken out and quickly shaken in a water bath at 37°C to melt them; the concentration of mononuclear lymphocytes is adjusted with GT‑T551 serum-free medium , and then add the mononuclear lymphocytes with adjusted concentration into the six-well plate at 2-4mL/well, adhere to the wall for 1-2 hours, and finally discard the upper layer of unattached cells in each well of the six-well plate; Add DC medium to each well of the orifice plate for culture, and continue to culture for 22-26 hours after adding the mixed factors that induce DC cells; and then culture for 22-26 hours after adding the factors that induce DC maturation. The DC cell preparation method provided by the present invention is very convenient, the time and scale of DC cell preparation can be well controlled, and the quality of the prepared DC cells is controllable, and the DC cell viability and purity are very high.

Description

technical field [0001] The invention relates to the field of cell preparation, in particular to a method for preparing DC cells. Background technique [0002] DC cells, the full name of dendritic cells (Dendritic Cells, DC), are professional antigen-presenting cells that have attracted much attention in recent years. They can efficiently ingest, process and present antigens, and initiate T-cell-mediated immune responses. Immature DCs have a strong ability to migrate, and mature DCs can effectively activate naive T cells, and are at the center of initiating, regulating, and maintaining immune responses. [0003] Conventional DC cell preparation is to isolate PBMC cells from fresh whole blood, and then directly enter the link of DC cell preparation. The acquisition of raw whole blood is accidental and limited, and the time and scale of DC preparation are not controllable. Contents of the invention [0004] The purpose of the present invention is to provide a method for prep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784
CPCC12N5/0639C12N2506/11
Inventor 王晓明刘倩苗嘉奕杨珺
Owner SHANGHAI ANGECON BIOTECH
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