Nov 03, 2022

Public workspacePassage cells

DOI
dx.doi.org/10.17504/protocols.io.n92ldpbd7l5b/v1
  • 1Institute of Tropical Disease, Airlangga University
Adita Ayu Permanasari
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DOI: dx.doi.org/10.17504/protocols.io.n92ldpbd7l5b/v1
Protocol Citation: Adita Ayu Permanasari 2022. Passage cells. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldpbd7l5b/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 06, 2022
Last Modified: November 03, 2022
Protocol Integer ID: 69611
Abstract
Passage Cells
Observe cells under microscope
Wipe outside surface of cell culture dish with ethanol cotton and bring it inside Biosafety Cabinet (BSC)
Bring medium, PBS, Trypsin-EDTA 2x to BSC
Bring autopipetter, disposable pipet (10ml, 2ml), one 15ml tube to BSC
Discard old medium
Rinse the bottom of dish with 10ml PBS
Add 1ml Trypsin-EDTA 2x to dish
Incubate the dish in CO2 Incubator 37oC 4min
Check under microscope (cell shape become rounded)
Neutralize trypsin-EDTA 2x by add 8ml medium
Suspend detached cells and transfer to 15ml tube
Centrifuge 1200 rpm, 4 min
Bring to BSC, discard medium
Add 20 ml DMEM and divided each 10 ml to dish (total 2 dish with @50% cells)