View source for Cultivation of HEK293T cells

== '''Materials''' ==
Medium: DMEM + 10 % calf serum (CS), PBS, Trypsin / EDTA

== '''Procedure:''' ==
* Remove growth medium with sterile pasteur pipette from dishes

* Wash 1x carefully with PBS, remove PBS

* 1 ml Trypsin/EDTA on cells, short incubation (max 2 min)
* If cells are detached, stop activity of trypsin with addition of 4 ml medium
* Transfer cell solution in falcon tube, centrifugation 800rpm, 3 min, room temperature
* Remove medium/ trypsin mixture, add new medium and mix
* ''Optional -'' For assays with specific cell numbers: 
** count cells in Neubauer counting chamber or use CasyTT: add 25 μl of cell solution in Casy tube
** add 5 ml of Casy Ton
** measure cell number
* For normal cultivation (two days) or transfection at next day: divide cells 1:5 in new dishes;  for cultivation over weekend (three days): divide cells 1:10 in new dishes


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