Signals from embryonic fibroblasts induce adult intestinal epithelial cells to form nestin-positive cells with proliferation and multilineage differentiation capacity in vitro

¹ In Vitro Differentiation Group, Institute of Plant Genetics and Crop Plant Research, Gatersleben, Germany
² Institute of Anatomy and Cell Biology, University of Halle-Wittenberg, Halle, Germany
³ Gerontology Research Center, National Institute on Aging, Natoinal Institutes of Health, Baltimore, Maryland
⁴ Institute of Reconstructive Neurobiology, University of Bonn, Bonn, Germany
⁵ Department of Pharmacology and Toxicology, Dresden University of Technology, Dresden, Germany
⁶ Institute of Reconstructive Neurobiology, University of Bonn, Bonn, Germany; Department of Epileptology, University of Bonn, Bonn, Germany
⁷ Department of Epileptology, University of Bonn, Bonn, Germany
⁸ Surgery University Clinics, Medical Faculty, University of Göttingen, Göttingen, Germany
⁹ DeveloGen AG Göttingen, Göttingen, Germany
¹⁰ Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, Bad Nauheim, Germany

^* To whom correspondence should be addressed. E-mail: wobusam{at}ipk-gatersleben.de.

	Abstract

The intestinal epithelium has one of the greatest regenerative capacities in the body; however, neither stem nor progenitor cells have been successfully cultivated from the intestine. In this study, we applied an 'artificial niche' of mouse embryonic fibroblasts to derive multipotent cells from the intestinal epithelium. Co-cultivation of adult mouse and human intestinal epithelium with fibroblast feeder cells led to the generation of a novel type of nestin-positive cells (INPs, intestinal epithelium-derived nestin-positive cells). Transcriptome analyses demonstrated that mouse embryonic fibroblasts expressed relatively high levels of Wnt/BMP transcripts, and the formation of INPs was specifically associated with an increase in Lef1, Wnt4, Wnt5a and Wnt/BMP-responsive factors, but a decrease of BMP4 transcript abundance. In vitro, INPs showed a high, but finite proliferative capacity and readily differentiated into cells expressing neural, pancreatic and hepatic transcripts and proteins, however these derivatives did not show functional properties. In vivo, INPs failed to form chimeras following injection into mouse blastocysts, but integrated into hippocampal brain slice cultures in situ. We conclude that the use of embryonic fibroblasts seems to reprogram adult intestinal epithelial cells by modulation of Wnt/BMP signalling to a cell type with a more primitive embryonic-like stage of development that has a high degree of flexibility and plasticity.