I am trying to establish intestinal absorption/permeability model using Caco2 cell line.
I seed Caco2 cells on 12 well permeable filters and grow them for 21 days for them to differentiate. I then measure TEER values of all wells including the blank (media only).
For the blank well, TEER is usually around 100 ohms while for the wells with cells its around 1100-1200 ohms, which seems to be quite high (most literature states around 500 ohms on day 21 to be optimal).
In order to solve this issue, I have tried using cell lines from different manufacturers, different FBS, cell media, seeding density and different passage numbers but the end result is the same.
Materials used:
1. Caco2 cells - ATCC and Leibniz
2. Voltohmmeter - Millicell ERS2 voltohmmeter
3. Cell detachment solution - TrpLE express enzyme (ThermoScientific)
4. Complete cell culture media - DMEM + 10% FBS + 1% non-essential amino acids + 1% Pen-strep
5. Permeable supports - 12-well insert PET (0.4 um) (CellQART)
Have you experienced similar issues? Ideas and suggestions are welcome :)
Thanks in advance!
Hi,
in my opinion your values are in the range of other studies. They have values up to 2000 ohm. But you are right, in the gut you find a value of 50-100 ohm. If you want to reduce the TEER, I would try another serum not FBS but human serum. Caco-2 cells have there origin in a human colorectal adenocarcinoma. So try to use human serum. A study for IPECs (intestinal porcine epithelial cells) exists, they have found high TEER values with FBS but low with pig serum...
PLoS ONE 8(11): e79643. doi:10.1371/journal.pone.0079643
best regards,
Constanze
Thank you very much for your input Constanze Nossol , I will try this out.
Hi Hitesh,
Like Constanze Nossol said, sounds like nothing is wrong.
Everything can change TEER, days since plating, media, FBS, Days since feeding, culture membrane etc....everything. Every lab will ave a different baseline resistance.
Just do your experiments carefully, do not pay too much attention to others, and i am sure your data will be different and believable.
MacG
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line