Is there anything that we have to add in media for the differentiation of HT-29 cells or HT-29 MTX specific cell line or any differentiation inducer is used for TEER estimation in HT-29 cells?? (because HT-29 cells are non-differentiating cell line). when I was growing the cells on transwell, the value of TEER was constantly, 150 from day 1 to 7 and after 7 days the cells are detached from the surface.
Hello, Deepti!
You look at this chapter:
https://www.ncbi.nlm.nih.gov/books/NBK500137/
Article Comparison of conventional and advanced in vitro models in t...
Cell differentiation and cell diversity in culture
Conventional culture lacks intense cell–cell interaction, signalling molecules and mechanical effects/dynamics. Furthermore, routine cytotoxicity testing is performed on subconfluent cells, a situation different from in vivo, where epithelial cells (intestinal, endothelial, respiratory, parenchymal cells of liver and kidney, etc.) are in direct intercellular contact and, with the exception of cells of the intestine, not constantly proliferating. Although cell lines show a decreased state of differentiation, they are preferred for basal toxicity screening because they possess all basal cellular functions and react in a more reproducible way than primary cells. To produce reliable data, cell lines have to be well characterized and to be routinely screened for bacterial contamination and for cross-contamination. Origin and use of cell lines in the different models mentioned in this review are provided in Table 2. To address cell-specific toxicity, cells need to express the specific phenotype and need to be treated in a specific way.
Different proliferation rates of the co-cultured cells may present a problem and limit the use of co-culture systems over longer time periods. In Caco-2/methotrexate-treated HT-29 (HT29-MTX) cell co-cultures, HT29 cells proliferate faster than Caco-2 cells, which results in the problem that the 9:1 ratio (Caco-2:HT29-MTX cells) rapidly changes over time. Instead of co-culturing cells in transwell systems, conditioned medium from one cell type can be added to another cell type at specific intervals. More recently, techniques were developed that enable a continuous exchange of media between cells. These techniques use small volumes and are summarized as microfluidics. Using continuous perfusion and chemical gradients, they represent better than conventional systems the microenvironment of cells in vivo. These systems are not yet established models for toxicity testing. A detailed description of the various microfluidics platforms is beyond the scope of this review and the reader is referred to reviews dedicated to this topic.
Article TEER Measurement Techniques for In Vitro Barrier Model Systems
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