Traditional Chinese medicine extract capable of improving CIK cell proliferation rate as well as preparation method and application of same
A cell proliferation and extract technology, applied in the direction of drug combination, pharmaceutical formulation, medical preparation containing active ingredients, etc., can solve the problems of low cytotoxicity, difficult cell source, clinical application limitation, etc. The effect of clear ingredients, good social and economic benefits
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Embodiment l
[0030] Example 1 A kind of formula and preparation method for preparing traditional Chinese medicine extract (SMH)
[0031] Take 5g of red ginseng, 5g of Ophiopogon japonicus, and 15g of astragalus. Heating purified water and decocting for extraction, the amount of water added is 10 times; decocting time is 0.5 hours. Collect the extract by filtration, concentrate the extract to obtain extract I; precipitate the above extract I with ethanol, filter, recover ethanol until it has no alcohol smell, and obtain refined extract II; enrich the refined extract II through a macroporous resin column , eluted with distilled water until colorless, eluted with ethanol, collected the eluate, concentrated, filtered through a 0.45 μm microporous membrane, and the filtrate was concentrated to 5 g crude drug / ml, boiled with activated carbon to absorb the heat source, removed carbon, and adjusted PH value, and filtered through a 0.22 μm microporous membrane to sterilize, subpackaged, and mad...
Embodiment 2
[0032] Example 2 Another formula and preparation method for preparing Chinese medicine extract (SMH)
[0033] Take 15g of red ginseng, 15g of Ophiopogon japonicus, and 30g of astragalus. Heating purified water and decocting for extraction, the amount of water added is 15 times respectively; the decocting time is 1 hour respectively. Collect the extract by filtration, concentrate the extract to obtain an extract; enrich the above extract through a macroporous resin column, elute with distilled water until it is colorless, elute with ethanol, collect the eluate, concentrate, and pass through a 0.45 μm micro Filtrate through a pore filter, concentrate the filtrate to 5g crude drug / ml, boil with activated carbon to absorb the heat source, remove carbon, adjust the pH value, filter through a 0.22μm microporous membrane to sterilize, sub-package, and make SMH Chinese medicine extract.
Embodiment 3
[0034] Example 3 A comparative experiment on the tumor cell killing ability of CIK cell culture added with traditional Chinese medicine extract (SMH) and conventional CIK cell culture after induction of proliferation and proliferation
[0035] (1) Co-stimulation culture with Chinese herbal extracts (SMH) and various cytokines
[0036] 30 mL of venous blood was collected in a vacuum blood collection tube containing anticoagulant, and peripheral blood mononuclear cells (PBMC) were separated with lymphocyte separation medium, washed twice, and then RPMI 1640 complete culture medium (containing inactivated 10% autologous plasma , 25 mmol / L HEPES, 100 U / ml penicillin, 100 U / ml streptomycin) to resuspend the cells and adjust the cell concentration to 2 × 10 6 / ml. The cells were equally divided into two groups, and the first group was cultured according to the conventional CIK cell culture method, that is, IFN-γ (1 000 U / ml) was added on the 0th day, and CO 2 After incubation in...
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