title: Reciprocal induction of simple organogenesis by mouse kidney progenitor cells in three-dimensional co-culture.
abstract: Kidney development is regulated by a coordinated reciprocal induction of metanephric mesenchymal (MM) and ureteric bud (UB) cells. Here, established MM and UB progenitor cell lines were recombined in three-dimensional Matrigel implants in SCID mice. Differentiation potential was examined for changes in phenotype, organization, and the presence of specialized proteins using immunofluorescence and bright-field and electron microscopy. Both cell types, when grown alone, did not develop into specialized structures. When combined, the cells organized into simple organoid structures of polarized epithelia with lumens surrounded by capillary-like structures. Tracker experiments indicated the UB cells formed the tubuloid structures, and the MM cells were the source of the capillary-like cells. The epithelial cells stained positive for pancytokeratin, the junctional complex protein ZO-1, collagen type IV, as well as UB and collecting duct markers, rearranged during transfection (RET), Dolichos biflorus lectin, EndoA cytokeratin, and aquaporin 2. The surrounding cells expressed α-smooth muscle actin, vimentin, platelet endothelial cell adhesion molecule 1 (PECAM), and aquaporin 1, a marker of vasculogenesis. The epithelium exhibited apical vacuoles, microvilli, junctional complexes, and linear basement membranes. Capillary-like structures showed endothelial features with occasional pericytes. UB cell epithelialization was augmented in the presence of MM cell-derived conditioned medium, glial-derived neurotrophic factor (GDNF), hepatocyte growth factor (HGF), or fibronectin. MM cells grown in the presence of UB-derived conditioned medium failed to undergo differentiation. However, UB cell-derived conditioned medium induced MM cell migration. These studies indicate that tubulogenesis and vasculogenesis can be partially recapitulated by recombining individual MM and UB cell lineages, providing a new model system to study organogenesis ex vivo.
authors.affiliation: Division of Nephrology, Department of Medicine, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900, USA.
authors.forename: Chakradhar
authors.initials: C
authors.lastname: Velagapudi
authors.forename: Rune-Par
authors.initials: RP
authors.lastname: Nilsson
authors.forename: Myung Ja
authors.initials: MJ
authors.lastname: Lee
authors.forename: Hannah S
authors.initials: HS
authors.lastname: Burns
authors.forename: Jill M
authors.initials: JM
authors.lastname: Ricono
authors.forename: Mazen
authors.initials: M
authors.lastname: Arar
authors.forename: Veronique L
authors.initials: VL
authors.lastname: Barnes
authors.forename: Hanna E
authors.initials: HE
authors.lastname: Abboud
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Barnes
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D003094:Collagen / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004338:Drug Combinations; D004847:Epithelial Cells / Q000648:ultrastructure; D061252:Feeder Cells / Q000502:physiology; D005353:Fibronectins / Q000378:metabolism; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000196:embryology* / Q000254:growth & development; D007797:Laminin / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051379:Mice; D016513:Mice, SCID; D009924:Organ Culture Techniques / Q000379:methods; D038081:Organogenesis / Q000502:physiology; D011509:Proteoglycans / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014513:Ureter / Q000166:cytology / Q000196:embryology* / Q000254:growth & development
chemical_list: D017077:Culture Media, Conditioned; D004338:Drug Combinations; D005353:Fibronectins; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D017228:Hepatocyte Growth Factor; D003094:Collagen