Culture medium for efficiently culturing umbilical cord blood CIK cells in vitro and culture method thereof

A technology of in vitro culture and culture medium, applied in the direction of blood/immune system cells, culture process, tissue culture, etc., to achieve the effect of simple operation, improved proliferation efficiency, and clear components

Active Publication Date: 2020-03-31
谭啸
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] With the traditional medium combination, the culture time needs 25-28 days before the number of cells reaches the plateau, and the number of cells and cytotoxic activity can reach the peak

Method used

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  • Culture medium for efficiently culturing umbilical cord blood CIK cells in vitro and culture method thereof
  • Culture medium for efficiently culturing umbilical cord blood CIK cells in vitro and culture method thereof
  • Culture medium for efficiently culturing umbilical cord blood CIK cells in vitro and culture method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A high-efficiency medium component for culturing CIK cells in vitro, adding cytokine combination A and traditional Chinese medicine extract B to AlyS505N-0 serum-free basal medium.

[0021] The specific components and contents of the medium are as follows:

[0022] No. 1: AlyS505N-0 serum-free basal medium + 800IU / ml IFN-γ + 100IU / ml IL-1α + 1200IU / ml IL-2 + 60μg / mlLymactin-T + 3μg / ml cordycepin;

[0023] No. 2: AlyS505N-0 serum-free basal medium + 800IU / ml IFN-γ + 200IU / ml IL-1α + 500IU / ml IL-2 + 20μg / ml Lymactin-T + 3μg / ml ginsenoside;

[0024] No. 3: AlyS505N-0 serum-free basal medium + 800IU / ml IFN-γ + 20IU / ml IL-1α + 2000IU / ml IL-2 + 100μg / ml Lymactin-T + 5μg / ml paclitaxel;

[0025] No. 4: AlyS505N-0 serum-free basal medium + 800IU / ml IFN-γ + 80IU / ml IL-1α + 1000IU / ml IL-2 + 80μg / ml Lymactin-T + 8μg / ml pachyman;

[0026] No. 5: AlyS505N-0 serum-free basal medium + 800IU / ml IFN-γ + 40IU / ml IL-1α + 800IU / ml IL-2 + 50μg / ml Lymactin-T + 4μg / ml astragaloside IV;

[0...

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Abstract

The invention discloses a culture medium for efficiently culturing umbilical cord blood CIK cells in vitro and a culture method thereof, which belong to the technical field of biology. The culture medium is prepared from the following components: an AlyS505N-0 serum-free basal culture medium, a cytokine composition A and a natural traditional Chinese medicine extract. The culture method comprisesthe following steps: collecting and separating umbilical cord blood mononuclear cells; resuspending cells, adding IFN-gamma for activation, incubating in a 6% CO2 incubator at 37 DEG C, adding the cytokine composition A for stimulation on the next day, supplementing the culture medium containing a traditional Chinese medicine extract B after 3-5 days, supplementing the culture medium containing the traditional Chinese medicine extract composition B every 2-3 days during culture, and co-culturing for 15-17 days to obtain a large number of CIK cells. According to the culture medium and the culture method thereof, a large number of CIK cells can be proliferated, and CD3+CD56+ double positive cells in the cells can reach 70% or above.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium and a culture method for efficiently culturing CIK cells in vitro. Background technique [0002] CIK, also known as multiple cytokine-induced killer cells, is a group of heterogeneous cells obtained by co-culturing human peripheral blood or umbilical cord blood mononuclear cells with various cytokines for a period of time in vitro. Because this kind of cells express both CD3+ and CD56+ membrane protein molecules, they are also called NK-like T lymphocytes, which have both the strong anti-tumor activity of T lymphocytes and the non-MHC-restricted tumor killing advantages of NK cells. [0003] CIK cells are extremely rare in normal human peripheral blood, only 1%-5%. After about 15 days of in vitro culture, the cells proliferate rapidly, and can be expanded by more than 1000 times compared with before culture. The effector cells express CD3+CD56+ at the sam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2500/30C12N2500/34C12N2500/40C12N2500/90C12N2501/2301C12N2501/2302C12N2501/999
Inventor 谭啸谭新民
Owner 谭啸
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