J774 Macrophage Cell Culturing
- J774 cell line, a murine macrophages cell line established from a tumor that arose in a female BALB/c mouse, were provided by Prof. Yi-Hsuan Lee from National Yang Ming University.
- View cultures using a microscopy and after the cultures was full of the dish, remove the spent medium.
- Wash the cell monolayer with PBS and remove it.
- Add medium(1ml per 25 cm2 of surface area) and scrape cells down.
- Ensure cells were detached by a microscopy.
- Centrifuge under 2000 rpm, 5minutes, 25°C.
- Remove the supernatant and add 5 ml medium to suspend cells.
- Count the cells by Bright Line Counting Chamber. Subject the cell diversity to 104/ml.
- Seed the cells for 104/well, 1ml/well in the 24-well plate.
- Until cells grow full of each well, do the coculturing experiment.
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