A drug and screening method for improving immunity under microgravity environment
A technology of immunity and microgravity, which is applied in the fields of biochemical equipment and methods, drug combinations, pharmaceutical formulations, etc., to achieve the effects of mild implementation conditions, rapid screening, and simple operation processes
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Embodiment 1
[0018] Example 1 Screening Polysaccharide Mixture Using Human Peripheral Blood Mononuclear Cells as Seed Cells
[0019] (1) Separation of human peripheral blood mononuclear cells by density gradient centrifugation
[0020] The heparinized anticoagulated blood was diluted one-fold with Hanks solution, spread lightly on Ficoll separation medium (specific gravity 1.077g / mL), and then centrifuged horizontally at 2000rpm for 30min. Mononuclear cells are suspended in the upper layer of the separation medium and appear as a white film. Carefully draw the mononuclear cell layer with a pipette, add Hanks solution and wash 3 times. Add RPMI-1640 medium containing 10% fetal bovine serum for routine culture.
[0021] (2) The prepared human peripheral blood mononuclear cell suspension (the total number of cells is 1 × 10 7 ) were cultured in a 50 mL rotary bioreactor, and phytolectin A (PHP-A) at a concentration of 20 μg / mL was added at the same time.
[0022] (3) Mix Astragalus Polysa...
Embodiment 2
[0024] Example 2 Screening Polysaccharide Mixture Using Mouse Bone Marrow Mononuclear Cells as Seed Cells
[0025] (1) Separation of mouse bone marrow mononuclear cells by density gradient centrifugation
[0026] Take the femur of the dead mouse, put the femur into sterile cold PBS; use a 1mL syringe to flush out the cells in the mouse femur until the femoral cavity is white; mix the cells well, use a 200-mesh filter to filter, collect and filter solution in a 10mL centrifuge tube, 1500rpm. Centrifuge for 5 minutes; discard the supernatant, add 8 mL of NH4Cl-Tris solution, suspend the cells, dissolve the red blood cells for about 6 minutes, centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, add 5mL sterile cold PBS to the centrifuge tube, and resuspend the cells; gently add the above cell suspension to 5mL Ficoll-paqul (1.086g / mL), centrifuge at 1500rpm for 30min; carefully take out the centrifuge tube, the solution Divided into three layers: transparent PBS laye...
Embodiment 3
[0030] Example 3 Screening Polysaccharide Mixture Using Mouse Spleen Lymphocytes as Seed Cells
[0031] (1) Separation of mouse spleen lymphocytes by density gradient centrifugation
[0032] The spleen tissue of the dead mouse was taken, placed in sterile cold PBS, then the spleen was cut into pieces, filtered through a 200-mesh filter, and the filtrate was collected in a 10mL centrifuge tube at 1500rpm. Centrifuge for 5 minutes; discard the supernatant, add 8 mL of NH4Cl-Tris solution, suspend the cells, dissolve the red blood cells for about 8 minutes, centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, add 5mL sterile cold PBS to the centrifuge tube, and resuspend the cells; gently add the above cell suspension to 5mL Ficoll (1.086g / mL), centrifuge at 2000rpm for 15min; carefully take out the centrifuge tube, and divide the solution into Three layers: from top to bottom are the transparent PBS layer, mononuclear cell layer, and RBS layer; use a thin pipette with...
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