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Knockout serum replacement for cell culture and preparation method thereof, knockout serum replacement composition for cell culture and cell culture medium

A technology of serum substitute and cell culture, applied in the field of serum substitute composition for cell culture, cell culture medium, serum substitute for cell culture and its preparation, can solve the problem of no serum-free medium, etc., to reduce the difference in results As well as the probability of infection by viruses and mycoplasma, high cell density and cell viability, and the effect of reducing dosage

Active Publication Date: 2019-07-09
河南普诺易生物制品研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows small amounts (less than 1 milliliter) of blood plasma from healthy individuals or patients with specific diseases to become suspended during experiments on these tiny particles called microorganisms. These microspheric organism carriers have been found that help keep them alive but they don't make any harm if their presence becomes too much. They also allow other living things like bacteria to stay within this space without being affected. By replacing only one part of the original solution containing all three components - Serum albumin, Plasminogen activator, and trypsins – we aimed at creating an improved method for producing proteins such as antibodies through cell cultivation techniques.

Problems solved by technology

This patented technical problem addressed in this patent relates to developing an effective method for growing and producing specific type of cells called HeLa's own stem cells while reducing costs associated with obtaining these cells from human tissue samples collected during surgery procedures.

Method used

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  • Knockout serum replacement for cell culture and preparation method thereof, knockout serum replacement composition for cell culture and cell culture medium
  • Knockout serum replacement for cell culture and preparation method thereof, knockout serum replacement composition for cell culture and cell culture medium
  • Knockout serum replacement for cell culture and preparation method thereof, knockout serum replacement composition for cell culture and cell culture medium

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preparation example Construction

[0037] The preparation method of the serum substitute for cell culture of the present invention comprises the following steps: dissolving each component in water, and then filtering and sterilizing.

[0038] Preferably, dissolving each component in water includes first dissolving in 450-550 mL of water, and then constant volume. Dissolving each component in water first is beneficial to the full dissolution and mixing of each component.

Embodiment 1

[0041] The serum substitute for cell culture in this example is composed of water and components with the following mass content: 18.5 mg of amino acid, 203 mg of protein, 62.9 mg of inorganic salt, and 1 mg of ethanolamine per liter of serum substitute for cell culture. Sodium pyruvate is 50 mg, ascorbyl phosphate is 1.25 mg, reduced glutathione is 0.5 mg, and insulin is 5 mg.

[0042] Amino acid consists of 2.25mg alanine, 3.5mg asparagine, 3.25mg aspartic acid, 3.5mg glutamic acid and 6mg proline; inorganic salt consists of 0.43mg ZnSO 4 ·7H 2 O, 0.00015mg ammonium metavanadate, 0.000625mg copper sulfate, 0.000025mg manganous chloride, 0.0025mg sodium selenite and 62.5mg Na 2 HPO 4 ·H 2 O composition; protein consists of 200mg albumin, 3mg transferrin.

[0043] The serum substitute for cell culture in this example is first prepared as a 10-fold concentrated solution, and then diluted 10-fold with water before use.

Embodiment 2

[0045] The serum substitute for cell culture in this example is composed of water and components with the following mass contents: 67 mg of amino acid, 815 mg of protein, 251.8 mg of inorganic salt, and 3.8 mg of ethanolamine in each liter of serum substitute for cell culture. Sodium pyruvate is 200 mg, ascorbyl phosphate is 5 mg, reduced glutathione is 2 mg, and insulin is 20 mg.

[0046] Amino acid consists of 9mg alanine, 15mg asparagine, 13mg aspartic acid, 15mg glutamic acid, 15mg proline; inorganic salt consists of 1.75mg ZnSO 4 ·7H 2 O, 0.0006mg ammonium metavanadate, 0.0025mg copper sulfate, 0.00010mg manganese chloride, 0.01mg sodium selenite, 250mg Na 2 HPO 4 ·H 2 O composition; protein consists of 800mg albumin, 15mg transferrin.

[0047] The serum substitute for cell culture in this example is first prepared as a 10-fold concentrated solution, and then diluted 10-fold with water before use.

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Abstract

The invention belongs to the technical field of cell culture, and particularly relates to a knockout serum replacement for cell culture and a preparation method thereof, a knockout serum replacement composition for cell culture and a cell culture medium. The knockout serum replacement for cell culture comprises, by weight, water, 18.5-67 parts of amino acid, 200-815 parts of protein, 62.5-252 parts of inorganic salt, 1.0-3.8 parts of ethanol amine, 50-200.0 parts of sodium pyruvate, 1.25-5.0 parts of ascorbyl phosphate, 0.5-2.0 parts of glutathione and 5.0-20 parts of insulin. The knockout serum replacement for cell culture can be added into any one cell culture medium, the adding amount of serum is reduced, and therefore, the cost is saved. The knockout serum replacement for cell cultureis suitable for protein recombination, vaccine production and other kinds of cell culture, and high in universality.

Description

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Claims

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Application Information

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Owner 河南普诺易生物制品研究院有限公司
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