title: Signals from embryonic fibroblasts induce adult intestinal epithelial cells to form nestin-positive cells with proliferation and multilineage differentiation capacity in vitro.
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. Cocultivation of adult mouse and human intestinal epithelium with fibroblast feeder cells led to the generation of a novel type of nestin-positive cells (intestinal epithelium-derived nestin-positive cells [INPs]). Transcriptome analyses demonstrated that mouse embryonic fibroblasts expressed relatively high levels of Wnt/bone morphogenetic protein (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 signaling to a cell type with a more primitive embryonic-like stage of development that has a high degree of flexibility and plasticity.
authors.affiliation: In Vitro Differentiation Group, Institute of Plant Genetics and Crop Plant Research, IPK, Corrensstrasse 3, D-06466 Gatersleben, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Wiese
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rolletschek
authors.forename: Gabriela
authors.initials: G
authors.lastname: Kania
authors.forename: Anne
authors.initials: A
authors.lastname: Navarrete-Santos
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Anisimov
authors.forename: Barbara
authors.initials: B
authors.lastname: Steinfarz
authors.forename: Kirill V
authors.initials: KV
authors.lastname: Tarasov
authors.forename: Sheryl A
authors.initials: SA
authors.lastname: Brugh
authors.forename: Ihor
authors.initials: I
authors.lastname: Zahanich
authors.forename: Christiane
authors.initials: C
authors.lastname: Rüschenschmidt
authors.forename: Heinz
authors.initials: H
authors.lastname: Beck
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Blyszczuk
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Czyz
authors.forename: Jürgen F
authors.initials: JF
authors.lastname: Heubach
authors.forename: Ursula
authors.initials: U
authors.lastname: Ravens
authors.forename: Olaf
authors.initials: O
authors.lastname: Horstmann
authors.forename: Luc
authors.initials: L
authors.lastname: St-Onge
authors.forename: Thomas
authors.initials: T
authors.lastname: Braun
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Boheler
authors.forename: Anna M
authors.initials: AM
authors.lastname: Wobus
mesh_terms: D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D004475:Ectoderm / Q000166:cytology; D004707:Endoderm / Q000166:cytology; D020895:Enterocytes / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D020869:Gene Expression Profiling; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D064231:Nestin; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015854:Up-Regulation / Q000235:genetics; D051153:Wnt Proteins / Q000235:genetics
chemical_list: D019485:Bone Morphogenetic Proteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D012333:RNA, Messenger; D051153:Wnt Proteins