Passage cells
Adita Ayu Permanasari
Published: 2022-11-03 DOI: 10.17504/protocols.io.n92ldpbd7l5b/v1
Abstract
Passage Cells
Steps
1.
Observe cells under microscope
2.
Wipe outside surface of cell culture dish with ethanol cotton and bring it inside Biosafety Cabinet (BSC)
3.
Bring medium, PBS, Trypsin-EDTA 2x to BSC
4.
Bring autopipetter, disposable pipet (10ml, 2ml), one 15ml tube to BSC
5.
Discard old medium
6.
Rinse the bottom of dish with 10ml PBS
7.
Add 1ml Trypsin-EDTA 2x to dish
8.
Incubate the dish in CO2 Incubator 37oC 4min
9.
Check under microscope (cell shape become rounded)
10.
Neutralize trypsin-EDTA 2x by add 8ml medium
11.
Suspend detached cells and transfer to 15ml tube
12.
Centrifuge 1200 rpm, 4 min
13.
Bring to BSC, discard medium
14.
Add 20 ml DMEM and divided each 10 ml to dish (total 2 dish with @50% cells)
