This is a cached page for the URL ( To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
Culturing HEK 293 Cells

Culturing HEK 293 Cells




Starting from a Kryotube:
Prewarm the medium at 37°C and fill two culture flasks (25 ml for 150 cm
2, 5 ml for 25 cm2). Rapidly thaw the cells (at 37°C) and distribute them in two concentrations in the flasks.
Change the medium after 12 hrs or once the cells have attached.

Splitting Cells:
Aspirate the medium from the flask. Wash the cells carefully with PBS to remove residual medium. Add 1-2ml of Trypsin Solution (equilibrated at RT) to the flask (150 cm
2) and incubate at 37°C until cells have detached (1-2 minutes). Prepare a new flask with fresh medium. Block trypsinization by adding a few ml of medium. Take a fraction of the cell solution and inocculate the new flask. Typically, when splitting confluent HEK 293 cells in a 1:10 ratio, confluency is reached again after 2-3 days.