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Use of chondroitin sulfate for expanding cik cells, preparing cik cell expansion reagents and coating culture vessels

A chondroitin sulfate, cell culture technology, applied in chondroitin sulfate, preparation of CIK cell expansion reagents and coated culture vessels, CIK cells, expansion of CIK cells field, can solve problems such as the application of chondroitin sulfate has not yet been seen, achieve The effect of improving tumoricidal activity and increasing expansion activity

Inactive Publication Date: 2019-07-02
辽宁惠泽康健生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no application of chondroitin sulfate in CIK cell culture expansion

Method used

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  • Use of chondroitin sulfate for expanding cik cells, preparing cik cell expansion reagents and coating culture vessels
  • Use of chondroitin sulfate for expanding cik cells, preparing cik cell expansion reagents and coating culture vessels
  • Use of chondroitin sulfate for expanding cik cells, preparing cik cell expansion reagents and coating culture vessels

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation and Characterization of High Molecular Weight Chondroitin Sulfate

[0024] 1. Preparation of chondroitin sulfate derived from chicken breast cartilage:

[0025] After the fascia is removed from the fresh chicken breast cartilage, it is dried at 55°C and crushed through a 40-mesh sieve to obtain cartilage powder. The cartilage powder is dissolved in a papain solution with a mass volume percentage of 8% (5g cartilage powder corresponds to 1L solution). Extract with papain at 55°C for 6 hours to obtain the enzyme extract, add trichloroacetic acid to a volume concentration of 10%, react for 1.5 hours, remove protein with trichloroacetic acid, and then filter and centrifuge at 4000-5000rpm for 8-12min. After the supernatant is concentrated, it is precipitated with ethanol (the volume of 95% ethanol is 5 times the volume of the supernatant), and the precipitate is collected by centrifugation at 4000-5000 rpm for 8-12 min. After the precipitation was re...

preparation example 1

[0027] Preparation Example 1: Dissolve the above-mentioned chondroitin sulfate raw materials with deionized water to prepare a chondroitin sulfate solution with a mass volume concentration of 3%, add malic acid to a concentration of 0.2mol / L, stir and degrade at 70°C for 10h, and use a sufficient amount of Calcium carbonate powder to terminate the reaction, adjust the pH to 6.5, add ethanol for precipitation (the volume of 95% ethanol is 8 times the volume of the degradation solution), centrifuge and dry at 55°C to obtain the high molecular weight chondroitin sulfate.

preparation example 2

[0028] Preparation Example 2: Dissolve the above-mentioned chondroitin sulfate raw materials with deionized water to prepare a chondroitin sulfate solution with a mass volume concentration of 2%, add malic acid to a concentration of 0.1mol / L, stir and degrade at 60°C for 12h, and use a sufficient amount of Calcium carbonate powder to terminate the reaction, adjust the pH to 6.5, add ethanol for precipitation (the volume of 95% ethanol is 8 times the volume of the degradation solution), centrifuge and dry at 55°C to obtain the high molecular weight chondroitin sulfate.

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Abstract

The invention discloses application of chondroitin sulfate in amplification of CIK cells, preparation of a CIK cell amplification reagent and coating of a culture vessel. The high-molecular-weight chondroitin sulfate is prepared from chondroitin sulfate sourced from animal breast cartilages through acid degradation, and the high molecular weight ranges from 14.2kD to 15.6kD. The high-molecular-weight chondroitin sulfate provided by the invention can obviously improve amplification vigor and cytotoxicity activity of the CIK cells and can be used for preparing commercial CIK cell culture medium and culture vessel.

Description

technical field [0001] The invention belongs to the field of cellular immunity and relates to CIK cells, in particular to the use of chondroitin sulfate for expanding CIK cells, preparing CIK cell expansion reagents and coating culture vessels. Background technique [0002] Chondroitin sulfate (CS) is widely found in various animal tissues, especially in cartilage and connective tissue. Since different organisms or different tissues of the same organism contain different types and amounts of CS, its structure and activity are complex and diverse. As a class of important biological macromolecules, CS has a wide range of biological activities, and has been widely used as a safe and effective health food and medicine in various countries in the world. A large number of studies have shown that CS has blood lipid-lowering, anti-atherosclerosis and anti-viral hepatitis activities, and has been used in the prevention and treatment of arthritis, nephritis, neuralgia, and migraine i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08C12N5/0783
CPCC08B37/0003C08B37/0069C12N5/0636C12N2501/90
Inventor 胡攀勇张钟祥胡珂
Owner 辽宁惠泽康健生物科技有限公司