title: Bronchoalveolar sublineage specification of pluripotent stem cells: effect of dexamethasone plus cAMP-elevating agents and keratinocyte growth factor.
abstract: Respiratory progenitors can be efficiently generated from pluripotent stem cells (PSCs). However, further targeted differentiation into bronchoalveolar sublineages is still in its infancy, and distinct specifying effects of key differentiation factors are not well explored. Focusing on airway epithelial Clara cell generation, we analyzed the effect of the glucocorticoid dexamethasone plus cAMP-elevating agents (DCI) on the differentiation of murine embryonic and induced pluripotent stem cells (iPSCs) into bronchoalveolar epithelial lineages, and whether keratinocyte growth factor (KGF) might further influence lineage decisions. We demonstrate that DCI strongly induce expression of the Clara cell marker Clara cell secretory protein (CCSP). While KGF synergistically supports the inducing effect of DCI on alveolar markers with increased expression of surfactant protein (SP)-C and SP-B, an inhibitory effect on CCSP expression was shown. In contrast, neither KGF nor DCI seem to have an inducing effect on ciliated cell markers. Furthermore, the use of iPSCs from transgenic mice with CCSP promoter-dependent lacZ expression or a knockin of a YFP reporter cassette in the CCSP locus enabled detection of derivatives with Clara cell typical features. Collectively, DCI was shown to support bronchoalveolar specification of mouse PSCs, in particular Clara-like cells, and KGF to inhibit bronchial epithelial differentiation. The targeted in vitro generation of Clara cells with their important function in airway protection and regeneration will enable the evaluation of innovative cellular therapies in animal models of lung diseases.
authors.affiliation: 1 Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School , Hannover, Germany .
authors.forename: Katherina
authors.initials: K
authors.lastname: Katsirntaki
authors.forename: Christina
authors.initials: C
authors.lastname: Mauritz
authors.forename: Ruth
authors.initials: R
authors.lastname: Olmer
authors.forename: Sabrina
authors.initials: S
authors.lastname: Schmeckebier
authors.forename: Malte
authors.initials: M
authors.lastname: Sgodda
authors.forename: Verena
authors.initials: V
authors.lastname: Puppe
authors.forename: Reto
authors.initials: R
authors.lastname: Eggenschwiler
authors.forename: Julia
authors.initials: J
authors.lastname: Duerr
authors.forename: Susanne C
authors.initials: SC
authors.lastname: Schubert
authors.forename: Andreas
authors.initials: A
authors.lastname: Schmiedl
authors.forename: Matthias
authors.initials: M
authors.lastname: Ochs
authors.forename: Tobias
authors.initials: T
authors.lastname: Cantz
authors.forename: Isabelle
authors.initials: I
authors.lastname: Salwig
authors.forename: Marten
authors.initials: M
authors.lastname: Szibor
authors.forename: Thomas
authors.initials: T
authors.lastname: Braun
authors.forename: Christian
authors.initials: C
authors.lastname: Rathert
authors.forename: Andreas
authors.initials: A
authors.lastname: Martens
authors.forename: Marcus A
authors.initials: MA
authors.lastname: Mall
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D003907:Dexamethasone / Q000008:administration & dosage*; D005240:Feasibility Studies; D051523:Fibroblast Growth Factor 7 / Q000008:administration & dosage*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods
chemical_list: D051523:Fibroblast Growth Factor 7; D003907:Dexamethasone; D000242:Cyclic AMP