Electro-transformation culture method of DC cells

By combining electroporation with the use of cytokines and nutrient solutions, the maturity and activity of DC cells were improved, solving the problems of insufficient DC cell activity and antigen expression in existing technologies, and achieving highly efficient DC cell culture.

CN122128237APending Publication Date: 2026-06-02GUANGDONG XIANKANGDA BIOTECH CO LTD
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Patent Information

Application Number
CN202610421435.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-04-01
Publication Date
2026-06-02

AI Technical Summary

Technical Problem

Existing technologies are insufficient for efficiently preparing mature dendritic cells (DCs) with high activity and high antigen expression. DCs prepared using existing technologies have low activity and low antigen expression, which cannot meet the needs of in vitro culture and clinical applications.

Method used

DC cell culture is performed by electroporation. This involves electroporation under specific conditions followed by culture, combined with the use of specific concentrations of cytokines and nutrient solutions, to improve the maturity and activity of DC cells.

Benefits of technology

Electroporation significantly improves the maturity and activity of DC cells, with high electroporation efficiency and shortened culture time, making it suitable for clinical application.

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Abstract

The invention discloses an electrotransfection culture method of DC (dendritic cell), which comprises the following steps: on the 0th day, inoculating a PBMC (peripheral blood mononuclear cell) suspension into a T175 culture flask, adding a first culture medium (comprising a basal culture medium, rhGM-CSF (recombinant human granulocyte-macrophage colony-stimulating factor), IL-4 (interleukin-4) and VC (vitamin C) components), and culturing in an incubator; on the third day, supplementing and adding the first culture medium, and continuously culturing; on the fifth day, absorbing and discarding the culture solution in the culture bottle, and supplementing and adding a second culture medium (including VC of the basic culture medium, rhGM-CSF, IL-4, IL-6, TNF-alpha and IL-1beta); on the seventh day, transferring the cells into an electrotransduction cup, adding an electrotransduction buffer solution, carrying out electrotransduction, adding a second culture medium, and continuously culturing; and on the tenth day, stopping culturing, collecting the cells, centrifuging, and removing supernatant to obtain the DC cells. The cells are cultured after being electrically transformed, so that the viability of the mature DC cells is greatly improved and reaches up to about 90%; the average fluorescence intensity (MFI) of the mature DC cells is greatly improved (as high as 10421), and the antigen expression quantity of the DC cells is good.
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