Spottl, Tanja and Hausmann, Martin and Gunckel, Manuela and Herfarth, Hans and Herlyn, Meenhard and Schoelmerich, Juergen and Rogler, Gerhard (2006) A new organotypic model to study cell interactions in the intestinal mucosa. EUROPEAN JOURNAL OF GASTROENTEROLOGY & HEPATOLOGY, 18 (8). pp. 901-909. ISSN 0954-691X,
Full text not available from this repository. (Request a copy)Abstract
Background Recently, we demonstrated that freshly elutriated monocytes differentiate into macrophages with a phenotype similar to that of intestinal macrophages in a three-dimensional model of intestinal epithelial cells. Here we describe a more organotypic model to study cell interactions in the intestinal mucosa. Methods Primary intestinal fibroblasts and freshly elutriated blood monocytes (ratio 1:1) were embedded in collagen type I gels and cultured for 5 days. At day 5, intestinal epithelial cells (HT-29) were seeded on top of the collagen gels. After another 7 days collagen gels were harvested and fixed for immunohistochemical analysis. Cryosections of the aggregates were prepared and staining for monocyte/macrophage markers and basement membrane compounds was performed. Cell interactions inside the aggregates were examined by electron microscopy. Results Intestinal fibroblasts contracted the collagen gels which formed stable three-dimensional aggregates within the first 5 days of culture. Intestinal epithelial cells formed a monolayer on top of the gels about 3 days after seeding. Intestinal fibroblasts were distributed randomly over the aggregate. Monocytes inside aggregates were localized in the vicinity to epithelial cells by positive staining for CD68. Typical monocyte/macrophage specific markers such as CD14, CD16, CD11b, CD11c and the co-stimulatory molecules CD80 and CD86 were down-regulated or not detectable on these cells after co-culture in three-dimensional aggregates. Omission of epithelial cells from the model was followed by impaired differentiation of intestinal macrophages. Conclusion In the three-dimensional organotypic cell culture model monocytes differentiate into intestinal-like macrophages when co-cultured with control intestinal fibroblasts and intestinal epithelial cells. Intestinal epithelial cells may be necessary for differentiation of intestinal macrophages.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | INFLAMMATORY-BOWEL-DISEASE; HUMAN SKIN EQUIVALENT; IN-VITRO MODEL; FACTOR-KAPPA-B; EPITHELIAL-CELLS; CULTURED SKIN; COLONIC MACROPHAGES; NODULAR CARCINOMAS; TISSUE CULTURE; DIFFERENTIATION; intestinal macrophages; cell differentiation; organotypic cell culture; intestinal mucosa |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 09 Feb 2021 05:49 |
| Last Modified: | 09 Feb 2021 05:49 |
| URI: | https://pred.uni-regensburg.de/id/eprint/34215 |
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