Publikationsdatenbank

Generation of 3D cell culture models (spheroids/organoids) to study interactions of zoonotic agents with the bovine intestinal mucosa (2019)

Art

Poster

Autoren

Haeger, Jan-Dirk

Berens, Christian

Sharbati, Soroush (WE 3)

Menge, Christian

Kongress

Zoonoses 2019 - International Symposium on Zoonoses Research

Berlin, 16. – 18.10.2019

Quelle

Zoonoses 2019 - International Symposium on Zoonoses Research : Book of Abstracts — International Symposium on Zoonoses Research (Hrsg.)

Berlin, 2019 — S. 150

Sprache

Englisch

Verweise

URL (Volltext): https://evis.events/event/79/attachments/23/154/Book_of_Abstracts_Zoonoses2019.pdf

Kontakt

Institut für Veterinär-Biochemie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62225
biochemie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

3D cell cultures resemble the histological structure of tissues more closely than monolayers.The aim of our studyis to establish 3D cultures which consist of various intestinal cell types isolated from cattle. Hanging drops (2 days) or matrigel (10 days) approaches were utilized to generate the 3D cultures. Primary cells (fibroblasts) and stable cell lines (FKD-R) were used as starting material, as well as primary colonic crypts. Different combinations of cells were employedin the hanging drops assays.We also approached the isolation of primary bovine intestinal crypt epithelial cells (pCEC) by use of enzymatic digestion.
When crypts were seeded in matrigel, they formed organoids (1000μm) which were demarked to the outside, as assessed by light microscopy. Hematoxylin-Eosin staining indicated that the organoids were made up of intestinal epithelium. In hanging drops, all cell combinations yielded cell spheroids. The spheroids/organoids survived harvest as they were mechanically stable, thereby providingproof for the formation of in vitro microtissues. The protocols for the continuous pCEC (2D) culture require optimization.
The established bovine intestinal colonic 3D-culture models will be employed to analyse the interplay of foodborne zoonotic agents transmitted from cattle to humans (e.g. Shiga toxin-producingEscherichia coli [STEC]).