Abstract
We examined an in vitro system to screen for diarrheagenic chemicals using an established intestinal cell line (T84 human colonic carcinoma). The cells were grown on Millicell-PCF (polycarbonate membrane) wells. The cells were seeded at approximately 5 × 106 cells/30mm well and incubated for 9–11 days in a 5% C02 incubator saturated with water at 37°C. The culture medium was a 1:1 mixture of Ham's F12 and Dulbecco's MEM with 5% fetal bovine serum and 25 pglml gentamicin sulfate. The well containing cells was removed from the incubator and mounted in a modified Ussing chamber for measurement of shortcircuit current (Isc). Chemical-induced increases in Psc are usually indicative of electrogenic epithelial Cl− secretion, which is associated with diarrheagenic effects in animals and humans. T84 cells grown on Millicell-PCF membrane responded with an increase in Isc after basolateral addition of the cholinergic (muscarznic) agonist carbachol, prostaglandin E2, 16,16-dimethylprostaglandin E2, and forskolin, while non-diarrheagenic prostaglandin D2 did not affect Isc. Based on our results, this in vitro system has the potential to be adapted as a rapid screen for detecting diarrheagenic chemicals.
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Abbreviations
- dmPGE2 :
-
16,16-dimethylPGE2
- EC50 :
-
50% of maximum effective concentration
- EDTA:
-
ethylenediaminetetraacetate
- ISC :
-
short-circuit current
- PGD2 :
-
prostaglandin D2
- PGE2 :
-
prostaglandin E2
- PD:
-
potential difference
- RT :
-
transepithelial resistance
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Oshiro, Y., Kachur, J.F., Balwierz, P.S. et al. An in vitro system to screen for diarrheagenic chemicals. Cell Biol Toxicol 9, 85–94 (1993). https://doi.org/10.1007/BF00755142
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DOI: https://doi.org/10.1007/BF00755142