In vitro culture method of human mature dendritic cells and application thereof

By combining density gradient centrifugation with a specific cytokine culture medium, peripheral blood mononuclear cells were efficiently induced to differentiate into high-purity, highly active mature dendritic cells. This solved the problems of limited expansion quantity and uneven maturity in existing technologies, enabling simple and low-cost preparation of dendritic cells suitable for tumor immunotherapy.

CN122214263APending Publication Date: 2026-06-16THE AFFILIATED HOSPITAL OF HANGZHOU NORMAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
THE AFFILIATED HOSPITAL OF HANGZHOU NORMAL UNIV
Filing Date
2026-03-24
Publication Date
2026-06-16

AI Technical Summary

Technical Problem

Existing methods for obtaining dendritic cells in vitro suffer from limitations such as limited cell expansion, uneven maturity, cumbersome operation, or high cost, which restrict their widespread application in clinical research and application.

Method used

Peripheral blood mononuclear cells were isolated using density gradient centrifugation and induced to differentiate into highly pure, highly active mature dendritic cells in vitro using specific cytokines and adjuvants. The process included Ficoll density gradient centrifugation, culture medium containing cytokines and adjuvants, and maturity was identified by flow cytometry.

Benefits of technology

It achieves the acquisition of high-purity, highly active mature dendritic cells, is simple to operate, and has controllable costs, making it suitable for basic scientific research and clinical treatment research, and possessing strong antigen presentation capabilities.

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Abstract

The application discloses an in-vitro culture method of human mature dendritic cells. Firstly, peripheral blood mononuclear cells are separated from human peripheral blood; then, monocytes are screened through adherent culture, and are induced to differentiate into immature dendritic cells in a first culture medium containing granulocyte-macrophage colony-stimulating factor and interleukin 4; finally, the immature dendritic cells are transferred into a second culture medium containing the same cell factors, antigen polypeptides and adjuvants, so that the immature dendritic cells are stimulated to further mature into mature dendritic cells loaded with antigens. The method has clear operation steps, low cost and good cell amplification effect. Through flow cytometry identification, the prepared mature dendritic cells have a purity of more than 97%, highly express mature markers such as CD11c, CD83, CD80 and CD86, have excellent antigen presentation function, and are suitable for preparing dendritic cell-based therapeutic vaccines or immunotherapy products.
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