Hello Stefanie,

after thawing the cells, we use our Caco-2 directly for experiments (but only for 10 passages) ...IPEC-1 and IPEC-J2 for 20 passages after thawing...

Best regards,

Constanze

Hi Constanze,

thank you for your answer.

Do you have any references for this? Why do you directly start and only use it for 20 passages?

All the best,

Stefanie

It is indeed known that cell passage is correlated with TEER values (see below).

From Srinivasan et al. "TEER measurement techniques for in vitro barrier model systems".

The study they mention in the first paragraph: Lu S., Gough A. W., Bobrowski W. F., et al. Transport Properties Are Not Altered across Caco-2 Cells with Heightened TEER Despite Underlying Physiological and Ultrastructural Changes. J. Pharm. Sci. 1996, 85, 270–273.

"The effect of passage number of Caco-2 cells on TEER measurements has been studied. TEER values were compared between the early passages of Caco-2 cells (Caco-2E, passages 35-47) and later passages of the cells (Caco-2L, passages 87-112). It was observed that the TEERs for Caco-2E cells ranged from 475 – 700 Ω.cm2 when compared to Caco-2L cells which ranged from 1100 – 1500 Ω.cm2. Transmission electron microscopy studies of these two cell populations revealed that regions of Caco-2L cells were composed of multiple cells layers rather than the monolayers observed in Caco-2E cells even though the epithelial cell height (barrier thickness) was not significantly different. Also, intercellular and intracellular lumina were observed in the Caco-2L cells, but not in Caco-2E cells. It was concluded that while intracellular lumina may not influence TEER values, the multiple cell layers and intercellular lamina may have significant impact on TEER measurements by changing the tightness and/or the length of the paracellular pathway.

Another study on the effect of passage number on TEER values has also been reported where Caco-2 cells from passage 20 through passage 109 were examined. It was observed that TEER values increased until passage 36, then the resistance was variable until passage 70, after which there was a decline in TEER value until passage 100."

Hello

After thawing Caco-2 cells needs to have 2 passages before starting an experiment

Caco-2 cells require specific culture conditions with a daily change of culture medium (DMEM 4,5g/L glucose; 1% AANE; 20% inactivated serum in 10% C02 incubator). A daily check of the cells is required.