Application of rapid Caco-2 cell culture system in the studies on the bacterial adhesion and transepithelial transport

• Autorzy: Olejnik A. , Marecik R. , Skrzypczak M. , Czaczyk K. , Grajek W.
• Języki publikacji: EN

Abstrakty

EN

The aim of the presented work was the functional comparison of conventional 21-day-old and modified 6-day-old Caco-2 cell cultures for their application in studies on the bacterial adhesion and transepithelial transport. In the short-term culture system, the cells were grown on polycarbonate filters in serum reduced medium supplemented with a combination of growth factors. Morphological comparison of two Caco-2 monolayers showed that the short-term culture had the similar ultrastructural characteristics of the long-term well-developed culture. The transepithelial electrical resistance measurement confirmed the high Caco-2 monolayer integrity in both cell culture systems. As a result, the modified 6-day-old Caco-2 culture could be applied in permeability and adhesion investigations. In the transport study, atrazine was used as a model molecule. The transport of atrazine was symmetric between the apical to basolateral and the basolateral to apical directions and independent of its concentration. No significant difference was found in atrazine permeability through conventional and modified Caco-2 cultures. In order to investigate the role of Caco-2 culture protocol might play in the adhesion of probiotic bacteria, three lactobacilli strains were used. The effect of Caco-2 culture modification on adhesion capacity of all tested bacteria was not observed. The proposed system of Caco-2 cell culture can be a considerable alternative to the conventional 21-day system.

Słowa kluczowe

EN

Caco-2 cell culture; application; probiotic adhesion; bacterial adhesion; transepithelial transport; food microbiology; biotechnology

• Wydawca: -
• Czasopismo: Polish Journal of Food and Nutrition Sciences
• Rocznik: 2008
• Tom: 58
• Numer: 3
• Opis fizyczny: p.365-371,fig.,ref.

Twórcy

autor

Olejnik A.

• Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznan, Poland

autor

Marecik R.

autor

Skrzypczak M.

autor

Czaczyk K.

autor

Grajek W.

Bibliografia

• 1. Anderle P., Niederer E., Rubas W., Hilgendorf C., Spahn-Langguth H., Wunderli-Allenspach H., Merkle H.P., Langguth P., Glycoprotein (P-gp) mediated efflux in Caco-2 cell monolayers: the influence of culturing conditions and drug exposure on P-gp expression levels. J. Pharm. Sci., 1998, 87, 757–762.
• 2. Blum S., Reniero R., Schiffrin E.J., Crittenden R., Mattila-Saldholm T., Ouwehand A.C., Salminen S., Wright A., Saarela M., Saxelin M., Collins K., Morelli L., Adhesion studies for probiotics: need for validation and refinement. Trends Food Sci. Technol., 1999, 10, 405–410.
• 3. Bradford M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 1976, 72, 248–254.
• 4. Braun A., Hammerle S., Suda B., Rothen-Rutishauser B., Gunthert M., Kramer S.D., Wunderli-Allenspach H., Cell cultures as tool in biopharmacy. Eur. J. Pharm. Sci., 2000, 11, S51–S61.
• 5. Coconnier M.H, Klaenhammer T.R., Kerneis S., Bernet M.F., Servin A.L., Protein-mediated adhesion of Lactobacillus acidiphilus BG2FO4 on human enterocyte and mucus-secreting cell lines in culture. Appl. Environ. Microbiol., 1992, 58, 2034–2039.
• 6. International Agency for Research on Cancer. Summaries and evaluations. Atrazine., 1999, 73, 59. Available at: [http://www.inchem.org/documents/iarc/vol73/73–03.html]
• 7. Jenkins N., Growth factors. 1991, in: Mammalian Cell Biotechnology. A Practical Approach (ed. M. Butler). Oxford University Press, pp. 39–55.
• 8. Kettles M.K., Browning S.R., Prince T.S., Horstman S.W., Triazine herbicide exposure and breast cancer incidence: an ecologic study of Kentucky counties. Environ. Health Perspect., 1997, 105, 1222–1227.
• 9. Korjamo T., Honkakoski P., Toppinen M-R., Niva S., Reinisalo M., Palmgren J.J., Monkkonen J., Absorption properties and P‑glycoprotein activity of modified Caco-2 cell lines. Eur. J. Pharm. Sci., 2005, 26, 266–279.
• 10. Lentz K.A., Hayashi J., Lucisano L.J., Polli J.E., Development of a more rapid, reduced serum culture system for Caco-2 monolayers and application to the biopharmaceutics classification system. Int. J. Pharmac., 2000, 200, 41–51.
• 11. Liang E., Chessic K., Yazdanian M., Evaluation of an accelerated Caco-2 cell permeability model. J. Pharm. Sci., 2000, 89, 336–345.
• 12. Meunier V., Bourrie M., Berger Y., Fabre G., The human intestinal epithelial cell line Caco-2; pharmacological and pharmacokinetic applications. Cell Biol. Toxicol., 1995, 11, 187–194.
• 13. Mills P.K., Correlation analysis of pesticide use data and cancer incidence rates in California counties. Arch. Environ. Health, 1998, 53, 410–413.
• 14. Olejnik A.M., Lewandowska M., Grajek W., Czaczyk K., New rapid method of Caco-2 cell differentiation. Pol. J. Food Nutr. Sci., 2003, 12/53, 60–64.
• 15. Pinto M., Robine-Leon S., Appay M.D., Kedinger M., Triadou N., Dussaulx E., Lacroix B., Simon-Assmann P., Hafen K., Fogh J., Zweibaum A., Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biol. Cell, 1983, 47, 323–330.
• 16. Pontier C., Pachot J., Botham R., Lenfant B., Arnaud P., HT29- MTX and Caco-2/TC7 monolayers as predictive models for human intestinal absorption: role of the mucus layer. J. Pharm. Sci., 2001, 90, 1608–1619.
• 17. Salminen S., Laine M., Wright A., Vuopio-Varkila J., Korhonen T., Mattila-Sandholm T., Development of selection criteria for probiotic strains to assess their potential in functional foods: a Nordic and European approach. Biosci. Microflora, 1996, 15, 61–67.
• 18. Sarem F., Sarem-Damerdji L.O., Nicolas J.P., Comparison of the adherence of three Lactobacillus strains to Caco-2 and Int-407 human intestinal cell lines. Lett. Appl. Microbiol., 1996, 22, 439–442.
• 19. Tavelin S., Grasjo J., Taipalensuu J., Ocklind G., Artursson P., Applications of epithelial cell culture in studies of drug transport. 2001, in: Methods in Molecular Biology, vol. 188 (ed. C. Wise). Humana Press Inc, Totowa, pp. 233–272.
• 20. Van Leeuven J.A., Waltner-Toews D., Abernathy T., Smith B., Shoukri M., Association between stomach cancer incidence and drinking water contamination with atrazine and nitrate in Ontario (Canada) agroecosystems. Int. J. Epidem., 1999, 28, 836–840.
• 21. Wunderli-Allenspach H., Methodologies in cell culture. 2000, in: Pharmacokinetic Optimization in Drug Research: Biological, Physico-chemical and Computational Strategies, vol. 1 (eds. B. Testa, H. Waterbeemd, G. Folkers, R. Guy). Wiley-VHCA, Zurich, pp. 218–252.
• 22. Yamashita S., Konishi K., Yamazaki Y., Taki Y., Sakane T., Sezaki H., Furuyama Y., New and better protocols for a short-term Caco-2 cell culture system. J. Pharm. Sci., 2002, 91, 669–679.