Adjustable controllable tumor living vaccine and its preparation method
A live vaccine and tumor technology, applied in the field of tumor live vaccine, tumor live vaccine and preparation thereof, can solve the problems of reducing vaccine antigenicity, transient, weak immune effect, 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 into 24-well plates, mrGM-CSF and mrIL-4 were added to each well, 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 wa...
Embodiment 2
[0038] 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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