Apr 03, 2025
Cell passage protocol for TR146 human buccal carcinoma cell line
- 1University of Minnesota
Protocol Citation: Boluwatife Olu Afolabi 2025. Cell passage protocol for TR146 human buccal carcinoma cell line. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvrkonplmk/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: April 03, 2025
Last Modified: April 03, 2025
Protocol Integer ID: 126034
Keywords: tr146, passage, splitting, cancer, culture
Funders Acknowledgements:
NIH/NIDCR
Grant ID: R21DE029337
Disclaimer
This protocol is provided as-is without any guarantees or warranties, expressed or implied, including but not limited to fitness for a particular purpose. Users are responsible for ensuring that the protocol is suitable for their specific experimental conditions and for complying with all relevant safety and ethical guidelines. The authors are not liable for any damages or issues arising from the use of this protocol.
Abstract
This protocol outlines the maintenance, subculturing, and cryopreservation of TR146 oral squamous carcinoma cells for use in cancer research
Materials
- DMEM/F12 Medium with 10% FBS
- Trypsin-EDTA (0.25%)
- DMSO (20% in Medium)
- PBS (Phosphate-Buffered Saline)
- 15 mL Conical Tubes
- Tissue Culture Flasks (T75, T50, T25)
- Transfer Pipettes
- Centrifuge
- 37°C Water Bath
- Personal Protective Equipment (PPE)
Safety warnings
Ensure a sterile environment to prevent cell contamination
Subculturing TR146 Cells
Subculturing TR146 Cells
10m
10m
Aspirate spent media using sterile vacuum system
Wash monolayer with 5 mL PBS at room temperature
Add 3 to 5 mL trypsin (37°C pre-warmed)
Incubate immediately for 5 min at 37°C (5% CO₂)
5m
Neutralize with 5 mL complete DMEM-F12
Centrifuge suspension: 300×g
5m
Remove supernatants via suctioning
Resuspend pellet in 3 mL fresh medium
Seeding Densities
Seeding Densities
| Vessel | Cell Suspension | Medium Volume | |
| T75 | 400 μL | 10 mL | |
| T25 | 200 μL | 5 mL |
Recommended seeding densities
Cryopreservation
Cryopreservation
Resuspend pellet from step 6 in 1.5 mL medium
Add 1.5 mL 20% DMSO-medium (keep ice-cold)
Aliquot 1 mL/vial (3 vials per T75)
Freeze at -80°C (overnight) → LN2 storage
Acknowledgements
Dr. Mark Herzberg
Dr. Chong Wang
Dr. Karen Johnstone