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文献和实验ISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTS
. It will be very acidic. After 3-4 days the culture will need splitting. Remove media and gently wash the monolayer with 2 X 10ml PBS. Add 2ml trypsin EDTA and split 1:4. After a further 2-4 days the culutre will be ready for freezing. The number obtained from each flask
density exceeds 1 x 106 cells/ml, dilute the cells to approximately 0.5 x 106 CD3 T cells/ml in Complete Medium. 7) Split the cultures to new 3L bags when needed. [Note: T cell growth typically slows as T cell concentrations increase
in the wells. 5. Load the plates into a centrifuge with a horizontal microtiter plate rotor and spin at 2600 x g for 40 minutes at 4°C. 6. Aspirate the supernatant from each well using the Immunowash plate washer. Settings for the depth
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