Antigen sensitized dendritic cell preparing method

Abstract

The invention relates to an antigen sensitized dendritic cell (DC) preparing method. On one hand, a culture medium for preparing antigen sensitized DCs is provided and is composed of a basal culture medium and other ingredients, the basal culture medium and the other ingredients can be conveniently mixed into the culture medium for culturing the DCs before using, the other ingredients comprise pharmaceutical grade human albumin, recombinant human insulin and the like, and the culture medium has the advantages of being definite in ingredient, free of ingredients of animal origin and the like; on the other hand, the invention provides a method for preparing the antigen sensitized DCs through the culture medium, and the method includes the steps that a single karyocyte is cultured and induced through the culture medium to obtain immaturate DC cells; strong immunogenicity tumor antigens are prepared through high hydrostatic pressure, astragalus polysaccharide is united to serve as an immunologic adjuvant, antigen sensitization is conducted on the immaturate DCs, and the antigen sensitized DCs are obtained. The preparing method has the advantages of being safe, efficient, easy to implement and the like. The DC vaccines for specific therapy can be provided for clinic through the method.

Description

technical field [0001] The invention relates to a method for preparing antigen-sensitized dendritic cells, belonging to the technical fields of cell engineering and biomedicine. Background technique [0002] Somatic cell therapy refers to the treatment method of using human autologous, allogeneic or heterogeneous (non-human) somatic cells, which are reinfused (or implanted) into the human body after extracorporeal manipulation. This in vitro operation includes cell passage, expansion, screening, and drugs or other treatments that can change the biological behavior of cells. The somatic cells after in vitro operation can be used for the treatment of diseases, and can also be used for the diagnosis and prevention of diseases. There are many different types of somatic cell therapy, including in vivo reinfusion of mononuclear cells activated in vitro, such as lymphokine activated killer cells (Lymphokine Activated Killer Cells, LAK), tumor infiltrating lymphocytes (Tumor Infiltr...

AI Technical Summary

Problems solved by technology

[0007] Although the serum-free medium has many advantages that the traditional serum-containing medium cannot match, it still has the following deficiencies to be improved: a. Cells are susceptible to certain mechanical and chemical factors in the serum-free medium. The preservation and application of the medium are not as convenient as the traditional synthetic medium; b. Strong pertinence, different types of cells, even different cell lines or cell strains have different requirements for the nutrients in the medium; c. Use serum-free medium for original d. Cell function may be changed to cause cell dysfunction; e. At present, most of the insulin and transferrin necessary for serum-free medium are still derived from animals, and the serum band has not been eliminated. potential safety hazards; f. higher cost

[0008] In order to improve and solve the above problems, researchers are constantly exploring and improving the formulation of serum-free medium for immune cells. Patent CN201110081988.5 provides a human-derived serum-free medium, which is composed of human serum albumin for treatment, human Composed of recombinant insulin, human transferrin, human cholesterol, human catalase, 2-mercaptoethanol, ascorbic acid, linoleic acid, ethanolamine, human vitronectin, L-glutamine, the added protein and lipid All are derived from human plasma, serum or tissue, and are pharmaceutical-grade or highly purified human-derived proteins or human recombinant proteins, which do not contain animal-derived components. Although this invention avoids the safety hazards of animal-derived substances, the human-derived protein Or human recombinant protein is expensive, which is not conducive to large-scale promotion and use; patent CN201310082166.8 provides a serum-free medium for the cultivation and proliferation of lymphocytes with clear chemical composition and no animal origin. The composition is complex and contains IMEM basal medium, L-glutamine, sodium bicarbonate, recombinant human insulin, human transferrin, human serum albumin, 2-mercaptoethanol, N-acetyl-cysteine, lipids, amino acids, microorganisms, Trace Minerals, Hydrocortisone, Ethanolamine, Non-Essential Amino Acids & Ferric Citrate

However, this product is only suitable for the culture and proliferation of lymphocytes, and the human recombinant protein or human protein used is expensive, which has great limitations in use

Images