Immune cell culture medium and culture method, as well as application of ginsenoside
A technology of immune cells and ginsenosides, applied in the application of ginsenosides, the field of immune cell culture medium, can solve the problems of poor immune cell activity and proliferation ability, etc.
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[0033] The immune cell culture medium provided by the invention includes ginsenoside, and the mass content of the ginsenoside in the serum-free basal medium is 4 mg / mL-10 mg / mL. In a specific embodiment of the present invention, the mass content of the ginsenosides in the serum-free basal medium can be 4 mg / mL, or 6 mg / mL, or 10 mg / mL; the present invention has no special restrictions on ginsenosides. Restrictions, the use of ginsenosides well known to those skilled in the art can be used, such as commercially available products can be used, or the preparation technical scheme for preparing ginsenosides well known to those skilled in the art can be used to prepare by itself. In the present invention, the preparation method of the ginsenoside preferably comprises the following steps:
[0034] extracting the coarse powder of ginseng stems and leaves, and filtering to obtain a primary filtrate;
[0035] The filtrate is removed from impurities, and then filtered to obtain a secon...
Embodiment 1
[0075] Take 40mL of peripheral blood, and use Ficoll separation method to obtain PBMC (mononuclear cells) 4×10 7 , divided into 10 groups, each group 4×10 6 Cells were inoculated in X-VIVO15 serum-free medium (purchased from LONZA Company), and ginsenosides were added thereto so that the concentrations of ginsenosides were 1 mg / mL, 2 mg / mL, 3 mg / mL, and 4 mg / mL, respectively. , 5mg / mL, 6mg / mL, 7mg / mL, 8mg / mL, 9mg / mL, 10mg / mL;
[0076] The inoculation density was 1×10 6 cells / mL; on the first day after inoculation, the cytokine INF-γ was added to a final concentration of 30 ng / mL; 80ng / mL;
[0077] Rehydration was performed every 3 days, and cytokine IL-2 was added according to the total volume after rehydration to a final concentration of 300 U / mL; after 14 days of culture, proliferated CIK cells were obtained.
[0078] The present invention counts the CIK cells after proliferation, and draws the curve of ginsenoside concentration and CIK cell proliferation multiple, such ...
Embodiment 2
[0080] Take 40mL of peripheral blood, and use Ficoll separation method to obtain PBMC (mononuclear cells) 4×10 7 , divided into 10 groups, each group 4×10 6 cells;
[0081] Groups 1 and 2 were used as control groups 1 and 2: groups 1 and 2 were induced to culture CIK cells using the serum-free medium in Comparative Example 1;
[0082] Groups 3 and 4 were used as control groups 3 and 4: Groups 3 and 4 used X-VIVO15 serum-free medium produced by LONZA Company in the United States to induce and culture CIK cells;
[0083] The 5th to 10th groups were used as the experimental groups of the present invention, wherein, the 5th and 6th groups were inoculated in X-VIVO15 serum-free medium, induced and cultured CIK cells, and the content of ginsenoside was 4 mg / mL;
[0084] Groups 7 and 8 were inoculated in X-VIVO15 serum-free medium to induce and culture CIK cells with a ginsenoside content of 10 mg / mL;
[0085] Groups 9 and 10 were inoculated in X-VIVO15 serum-free medium to induce a...
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