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A technology of serum-free medium and basal medium, applied in the field of culture medium, can solve the problems of low T cell culture medium culture efficiency, potential safety hazards, animal serum protein residues, etc.
Inactive Publication Date: 2018-10-12
安徽中盛溯源生物科技有限公司
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Problems solved by technology
These culture media are more or less added with animal-derived or human blood-derived components. Although there are strict testing and virus inactivation treatments, there are still certain potential safety hazards.
[0005] Therefore, the current T cell culture medium has major problems such as low culture efficiency, low cell activity, follow-up test interference, and animal serum protein residues.
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Embodiment 1
[0048] The T cell serum-free medium in this embodiment includes basal medium and other additives, wherein the basal medium is mixed with DMEM, F12, and IMDM at a volume ratio of 3:3:1, and other additives are added to the basal medium Components, so that each liter of serum-free medium contains ethanolamine 0.1mmol, ketone sulfate 1μg, ferric nitrate 0.5mg, zinc sulfate 0.3mg, sodium selenite 20μg, sodium pyruvate 150mg, insulin 10mg, recombinant human transferrin 20mg, Glutamine 5ml, recombinant human serum albumin 0.5g, thioglycerol 20μmol, L-vitamin C 20mg, interleukin-2 5×10 4 IU.
Embodiment 2
[0050] The serum-free medium for T cells in this embodiment includes basal medium and other additives, wherein the basal medium is mixed with RPMI-1640 and IMDM at a volume ratio of 1:1, and other additives are added to the basal medium, So that each liter of serum-free medium contains ethanolamine 0.05mmol, ketone sulfate 1.5μg, iron nitrate 1mg, zinc sulfate 0.3mg, sodium selenite 15μg, sodium pyruvate 110mg, insulin 15mg, recombinant human transferrin 50mg, glutamine Amide 25ml, recombinant human serum albumin 5g, thioglycerol 60μmol, L-vitamin C 50mg, interleukin-2 1.5×10 5 IU.
Embodiment 3
[0052] The T cell serum-free medium in this embodiment includes basal medium and other additives, wherein the basal medium is mixed with DMEM, F12, and IMDM at a volume ratio of 3:3:1, and other additives are added to the basal medium Components, so that each liter of serum-free medium contains ethanolamine 0.1mmol, ketone sulfate 0.5μg, ferric nitrate 0.6mg, zinc sulfate 0.8mg, sodium selenite 1.8μg, sodium pyruvate 70mg, insulin 0.5mg, recombinant human trans Ferritin 6mg, glutamine 5ml, recombinant human serum albumin 0.5g, thioglycerol 65μmol, L-vitamin C 30mg, interleukin-2 5×10 4 IU.
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Abstract
The invention belongs to the field of culture mediums, and in particular relates to a T cell serum-free medium and a using method thereof. The T cell serum-free medium consists of a basal culture medium applied to growth culture of cells, as well as the following addition ingredients: ethanolamine, copper sulfate, ferric nitrate, zinc sulfate, sodium selenite, sodium pyruvate, insulin, transferrin, glutamine, serum albumin, thioglycerol and L-vitamin C. The T cell serum-free medium provided by the invention breaks through shortcomings of the prior art, and according to the T cell serum-free medium, the entire medium system is free from the introduction of non-human proteins, and the T cells in the blood sample, which is safe and reliable in source, undergo efficient selective amplification; and therefore, higher clinical application and scientific research values are achieved.
Description
technical field [0001] The invention belongs to the field of culture medium, and in particular relates to a T cell serum-free culture medium and a use method thereof. Background technique [0002] T cells are one of the main components of lymphocytes. They have a variety of biological functions, such as directly killing target cells, assisting other lymphocytes to function, responding to specific antigens or mitogens, and producing cytokines. One of the body's main immune cells that fight off infection and prevent tumors from forming. Enrichment, activation, and expansion of such immune cells in vitro can be used for reinfusion to treat various diseases, including malignant tumors, infections, autoimmune diseases, etc. In this process, it is necessary to use a T cell that can support the survival of T cells in vitro and amplified media. [0003] At present, RPMI-1640 or DMEM medium is mostly used for T cell culture, but the nutrients required for the survival and expansion...
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