Jan 14, 2026
Cell culture
- Yongjia Duan1
- 1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA
Protocol Citation: Yongjia Duan 2026. Cell culture. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6ex2zgqe/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: January 14, 2026
Last Modified: January 14, 2026
Protocol Integer ID: 238648
Keywords: ASAPCRN, cell culture cell culture, cell
Abstract
Cell culture
Troubleshooting
HEK293-Lenti-X, Stable HeLa cell lines
These cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco)
supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS, Gibco)
and 1x anti-anti (Gibco).
Cells were routinely passaged every 2-3 days when they reached 80-90% confluence and maintained at 37℃ in 5% CO2
HEK293F GnTi cells
Cells were maintained in FreeStyle‱ 293 Expression Medium (Gibco) supplemented
with 1% FBS and 1x anti-anti.
Cells were routinely passaged every 2-3 days, maintained at 37℃ in 5% CO2, and Incubated with shaking at 100 rpm.
ARPE‐19 cells
Cells were maintained in 1:1 DMEM:F‐12 media (Thermo Scientific)
supplemented with 10% (v/v) FBS, 2 mM L‐glutamine, 100 U/ml penicillin and
0.1 mg/ml streptomycin
Cells were routinely passaged every 2-3 days when they reached 80-90% confluence and maintained at 37℃ in 5% CO2
293FT cells
Cells were cultured in DMEM with supplemented with 10% (v/v) FBS,
2 mM L‐glutamine, 100 U/ml penicillin and 0.1 mg/ml streptomycin
Cells were routinely passaged every 2-3 days when they reached 80-90% confluence and maintained at 37℃ in 5% CO2
All cell lines were confirmed mycoplasma negative using MycoAlert Detection kit every 1 month.