Serum-free medium suitable for culturing gamma delta T cells

Abstract

The invention discloses a serum-free medium suitable for culturing gamma delta T cells. The serum-free medium consists of a basal culture medium component I, a gamma delta T cell specific culture component II and water. Experiments show that by adopting the serum-free medium suitable for culturing the gamma delta T cells, the gamma delta T cells obtained is greater in quantity, higher in purity and better in killing effect. The serum-free medium suitable disclosed by the invention has the advantages that 1) the risk of animal components which are used in animal serums in the cell culture with the gamma delta T cells on patients with cell therapy and uncertain impact of uncertain components in the animal serums on the result of the cell culture are avoided; 2) the yield of the gamma delta T cells is greatly improved; and 3) the final density of the gamma delta T cell culture is increased so as to lower the cost of cell culture.

Description

technical field [0001] The invention relates to a serum-free medium suitable for culturing γδT cells. Background technique [0002] γδT cells were discovered in the mid-1980s. There are two main subtypes: Vγ9 / Vδ2 and Vδ1. γδT only accounts for 0.5% to 5% of peripheral blood T lymphocytes, of which more than 90% are Vγ9 / Vδ2 cells . Vδ1 is mainly distributed in epithelial tissues, such as intestine, spleen, skin, small intestine, esophagus, lung and reproductive organs. [0003] γδT cells have the characteristic of rapid response in vivo. When stimulated, it will release pro-inflammatory cytokines before αβ-T cells, such as IFN-γ and TNF-α; it has high lethality to tumor cells, and can recognize MICA / B on the surface of tumor cells through the cell membrane surface molecule NKG2D and DNAM-1 and other membrane proteins to initiate a variety of ways to kill tumor cells, such as the release of perforin, granzyme, FasL, Trail, etc.; γδT cells can also activate the tumor cytotox...

AI Technical Summary

Problems solved by technology

However, since animal serum contains animal ingredients, if it is used in a clinical cell therapy system, it may cause some potential risks. This is mainly due to the complex composition of animal serum. Although it contains many beneficial ingredients for cells, there are also many unfavorable cells. Growth factors exist: 1) Animal serum contains some substances that are toxic to cells, such as polyamine oxidase, which can react with polyamines from highly reproductive cells (such as spermine, spermidine) to form cytotoxic polyamines. Spermine, complement, antibodies, bacterial toxins, etc. will all affect cell growth and even cause cell death; 2) Individual animals are different, and serum origin and batch numbers are different, and the quality of each batch varies greatly, and its components cannot be kept consistent; 3) The samples may contain Mycoplasma and viruses may have potential effects on cells, which may lead to experimental failure or unreliable experimental results; 4) In large-scale production, the source of serum is becoming more and more difficult and expensive, which constitutes the main part of the production cost of animal cell culture one

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