Lymphocyte cultivation liquid and method and application

A technology of lymphocytes and culture medium, applied in the biological field, can solve the problems of providing sufficient energy for culture, affecting cell expansion and passage, etc., and achieve the effects of promoting expansion, long passage time, and high anti-tumor activity

Active Publication Date: 2007-09-19
THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its energy substance is only glutamine, so there is also the problem that it cannot provide enough energy for cell culture, thus affecting cell expansion and passage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The composition and consumption of the culture fluid of the present embodiment are shown in Table 1

[0043] components

Dosage

Basic culture medium RPMI-1640

10.4g / L

Albumin

10wt%

L-Glutamine

0.3g / L

glucose

2g / L

sodium bicarbonate

2g / L

HEPES

6g / L

ATP

10mg / L

coenzyme A

40U / L

[0044] The preparation method of the above-mentioned culture solution is as follows: add 10.4g basal culture solution RPMI-1640 (one bag), 10wt% albumin, 0.3g L-glutamine, 2g glucose, 2g sodium bicarbonate, 6gHEPES. At the same time, 40,000 U of gentamicin sulfate and 10 mg of fluconazole can also be added. Stir to dissolve it completely, filter and sterilize through a microporous membrane, and store at -20°C. Add 10 mg of ATP and 40 U of coenzyme A before use.

[0045] The above-mentioned pack of basal culture medium RPMI-1640 contains 10mmol / L HEPES, 0.02mmol / L 2-mercaptoethanol, 100U / ml ...

Embodiment 2～ Embodiment 5

[0064] The composition and consumption of the culture solution of embodiment 2~embodiment 5 are shown in table 4

[0065] components

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PUM

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Abstract

The invention discloses a lymphocyte culture fluid and culture method thereof and application therefor, the culture fluid is adding ATP and coenzyme A in completely culture fluid containing basic culture fluid RPMI-1640, albumin, L-glutamine, glucose, baking soda and HEPES. The culture method includes steps: (1) preparing culture fluid; (2) suspending cell in culture fluid prepared by step (1); (3) activating cell; (4) washing activated cell by the culture fluid prepared by step (1); (5) collecting cell. The culture fluid of the invention cultures cell with higher amplification times and longer generation time.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a lymphocyte culture medium and a method and application for culturing lymphocytes. Background technique [0002] Lymphocytes are a kind of immunologically active cells with important uses. By extracting lymphocytes from cancer patients and inducing culture in vitro, a large number of active cells with anti-tumor cell ability can be obtained. [0003] Cytokine-induced killer cells, referred to as CIK cells. It is a group of heterogeneous cells obtained after autologous peripheral blood mononuclear cells are co-cultured with various cytokines for a period of time in vitro. After a large amount of such cells are reinfused, it can provide tumor cell killing activity and make the body Immunity is enhanced, thereby exerting a dual effect on tumor therapy. Since CIK cells are activated autologous cells, they are safe and have no obvious side effects. Currently, CIK cells are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/0783
Inventor 蒋敬庭
Owner THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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