Nov 02, 2023

Public workspaceCell culture

DOI
dx.doi.org/10.17504/protocols.io.36wgq3bk5lk5/v1
  • 1University of Cambridge
Alexander Röntgen
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DOI: dx.doi.org/10.17504/protocols.io.36wgq3bk5lk5/v1
Protocol Citation: Alexander Röntgen 2023. Cell culture. protocols.io https://dx.doi.org/10.17504/protocols.io.36wgq3bk5lk5/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: November 02, 2023
Last Modified: November 02, 2023
Protocol Integer ID: 90290
Abstract
Protocol to culture SH-SY5Y cells.
Materials
  • Medium: DMEM/F-12, GlutaMAXTM (ThermoFisher Scientific) + 10% Fetal Bovine Serum (GibcoTM)
  • Trypsin-EDTA (0.25%) (ThermoFisher Scientific)
  • PBS, pH 7.4 (ThermoFisher Scientific)
Cell culture
Cell culture
Culture human SH-SH5Y cells at 37 °C and 5% CO2
Split cells at 80–90% confluency

Discard medium, rinse with PBS
Incubate cells with Trypsin-EDTA for 5 min at 37 °C and 5% CO2
Add 9 mL medium to neutralise Trypsin-EDTA
Centrifuge at 300 g, 5 min, RT
Discard medium
Resuspend in 10 mL medium
Split at desired ratio into fresh medium