Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Zegay, S.
Influence of Herbal Substances on the Intestinal Epithelium and its
Microbiota in an Air Liquid Interface Culture
Humanmedizin; [ Diplomarbeit ] Medizinische Universität Graz; 2022. pp. 47
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Brislinger Dagmar
- Moissl-Eichinger Christine
- Altmetrics:
- Abstract:
- Microbiota have been found to metabolize substances in different ways and can sometimes be the key factor of the therapeutic effectiveness of a substance. Run Chang Wan (RCW) is a traditional Chinese herbal medicine often used to relieve constipation, but the certain mechanisms of action are still unclear. Conventional cell cultures do not resemble the in vivo conditions, making it impossible to observe if there are relevant interactions between an agent, differentiated epithelial cells and microbiota. We tried to overcome this problem by combining microbiota within an Air Liquid Interface culture of colonic cells to resemble an environment as near as possible to in vivo conditions. The Air Liquid Interface is a cell culturing method to support the differentiation of epithelial cells into a pseudostratified ciliated epithelium with mucus producing goblet cells. Cells are cultured in inserts on a polycarbonate membrane with pores of 0.4 µm so the cells are exposed to air on the apical side and in contact with medium from the basal side. Two colon adenocarcinoma cell lines were cultured, CaCo2 and HT29, to form a colonic epithelium. The microbiota were applied as a 10% suspension in anaerobe medium and incubated with different concentrations of RCW. These conditions lead to cell damage as RNA measurements showed. Also, most staining procedures were not successful, probably due to a lack of vivid cells to adhere. Only the cells without RCW but with microbiota survived with visible signs of cell damage. Further studies need to be conducted to find successful methods to combine ALI cultured colon cells with microbiota.