Hello everyone,

I am encountering an issue with my Caco-2 cell cultures where the TEER values are not increasing and have remained around 40 ohms since day 3 post-seeding. Here's a summary of my protocol and observations:

• Plate Type: Millicell®-24 plates with an active membrane area of 0.7 cm² (1 µm; PET).
• Seeding Density: 60,000 cells/well, as recommended by Millipore.
• Culture Medium: DMEM (high glucose) + 10% FBS + 1% P/S.
• TEER Measurement: Using an older voltohmmeter, which reads slightly below 100 ohms for empty wells (suggesting it is functional).
• Observation: My Caco-2 cells seem to be growing unusually fast. They originally grew in RPMI medium, taking 4 days to reach confluency at a 1:10 split ratio. After switching to DMEM (high glucose), they reach confluency within 3 days, even at a 1:50 split ratio.

I have considered several potential issues:

Could the unusually rapid growth rate in DMEM (high glucose) affect the TEER values? Should I adjust the seeding density despite following the recommended protocol? Are there any other factors I might be overlooking that could explain the persistently low TEER values?

Any advice or insights from those with experience in Caco-2 culture and TEER measurements would be greatly appreciated.

Thank you!

#Caco-2 #TEER #Transwell