Adjustable controllable tumor eiving vacine and its preparation method
A technology of live vaccines and tumors, applied in the field of live tumor vaccines, live tumor vaccines and their preparation, can solve the problems of weak and short-lived immune effects, reducing vaccine antigenicity, etc.
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Embodiment 1
[0030] Mononuclear cells were isolated from the femoral bone marrow of F344 rats, red blood cells were dissolved in double distilled water, inoculated in 24-well plates, mrGM-CSF and mrIL-4 were added to each well, and half of the medium was changed every other day, and cultured for 8-10 days, each Rats can obtain dendritic cells 5~10×10 7 ;Typical colony growth of dendritic cells was observed under an inverted microscope; Fluorescence-labeled antibody flow cytometry was used for dendritic cell phenotype identification, and the results showed that the cell surface antigens OX6, B7-2, OX62 and ICAM were cultured for 6 days -1 were 20.89%, 22.68%, 21.93% and 13.74%, respectively; increased to 63.46%, 52.79%, 75.35% and 30.43% on day 10. The dendritic cells of F344 rats cultured on the 6th day can slightly stimulate the proliferation of T cells, with a stimulation index of 1.21±0.23; while the ability of the dendritic cells cultured on the 9th day to promote T cell proliferation ...
Embodiment 2
[0047] Mononuclear cells were isolated from the femoral bone marrow of F344 rats, red blood cells were dissolved in double distilled water, inoculated in 24-well plates, mrGM-CSF and mrIL-4 were added to each well, and half of the medium was changed every other day, and cultured for 8-10 days, each Rats can obtain dendritic cells 5~10×10 7 ; Typical dendritic cells grew into colonies under an inverted microscope; Gimsa staining, immunohistochemistry, scanning, and transmission electron microscopy showed typical dendritic cell morphology. Fluorescence-labeled antibody flow cytometry was used to identify the phenotype of dendritic cells. The results showed that the surface antigens of dendritic cells were lowly expressed when they were immature: 45.22% for OX6 (MHC class II), and 32.76% for B7-2; high expression when mature OX6 is 89.21%, and B7-2 is 56.33%. The suicide gene CD was transfected into the F344 rat ovarian cancer cell line NUTU-19 using adenovirus as a carrier, and...
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