Cell culture medium composition containing resveratrol and silk sericin
A technology of resveratrol and sericin, which is applied in the direction of cell culture medium, cell culture active agent, animal cells, etc., to achieve the effect of increasing expression level, improving in vitro amplification ability, and enhancing homing ability
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Embodiment 1
[0051] Preparation of stock solutions of resveratrol and sericin
[0052] Resveratrol powder (purchased from Sigma, USA) was dissolved in cell culture-grade DMSO to prepare a storage solution with a final concentration of 5 mM. The solution was sterilized by filtering through a 0.22 μm filter, and divided into 1 ml / bottle to avoid The light is stored at -80 degrees.
[0053] Silk sericin dry powder (purchased from Wako Company, Japan) was dissolved in PBS with a pH of 7.4 and prepared as a 10 mg / ml storage solution. The solution was sterilized by filtering through a 0.22 μm filter, divided into 1 ml / cartridge, and frozen in Save at -80 degrees.
[0054] When in use, take out the stock solutions of the two additives and melt them quickly at room temperature;
[0055] Mix with the culture medium according to the following ratio
[0056] Take 2ml of resveratrol storage solution, 1ml of sericin storage solution, 97ml of SCGM medium,
[0057] Mix well at room temperature, that ...
Embodiment 2
[0077] Example 2. Resveratrol combined with sericin to evaluate the proliferation ability of NK cells in vitro. Collect 30ml of peripheral blood from healthy people. According to the method reported in the literature, use human mononuclear cell separation medium (Beijing Dongfang Huahui Biopharmaceutical Co., Ltd. Technology Co., Ltd., Cat.No.25610) for the separation of PBMC. The separated PBMCs were counted by trypan blue exclusion method, the cell number was adjusted to 0.5x106 cells / ml, and resuspended with D-PBS.
[0078] According to the grouping method of the experimental design in Table 1., NK cells were cultured in vitro using different culture recipes. SCGM medium was purchased from CellGenix, humanized CD3 monoclonal antibody OKT-3 was purchased from Takara, recombinant human interleukin- 2 (IL-2) was purchased from Beijing Shuanglu Pharmaceutical.
[0079] The cultivation system is respectively,
[0080] Blank SCGM medium,
[0081] SCGM medium containing resvera...
Embodiment 3
[0089] Example 3. Effects of Resveratrol and Sericin on the Expression Levels of Different Molecular Receptors on the Surface of NK Cells
[0090] On the 14th day of culture, different groups of NK cells were collected, and the representative activating receptors, inhibitory receptors, chemotactic receptors and cytokine receptors expressed on the surface of NK were analyzed by flow cytometry. analyze. The operation process is briefly described as follows: the number of cells in each sample for flow cytometry is not less than 10 6 cell. The sample was centrifuged at 350g for 10 minutes at room temperature, the supernatant of the culture medium was discarded, and resuspended in cell preservation solution with a final volume of 200 μl. Add different specific antibodies according to the detected molecules, the detected molecules and the calculation method of the expression fold change of the corresponding molecules are shown in Table 2 below.
[0091] Table 2. NK cell surface m...
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