title: Enhanced lung epithelial specification of human induced pluripotent stem cells on decellularized lung matrix.
abstract: BACKGROUND
  Whole-lung scaffolds can be created by perfusion decellularization of cadaveric donor lungs. The resulting matrices can then be recellularized to regenerate functional organs. This study evaluated the capacity of acellular lung scaffolds to support recellularization with lung progenitors derived from human induced pluripotent stem cells (iPSCs).
  
  
  METHODS
  Whole rat and human lungs were decellularized by constant-pressure perfusion with 0.1% sodium dodecyl sulfate solution. Resulting lung scaffolds were cryosectioned into slices or left intact. Human iPSCs were differentiated to definitive endoderm, anteriorized to a foregut fate, and then ventralized to a population expressing NK2 homeobox 1 (Nkx2.1). Cells were seeded onto slices and whole lungs, which were maintained under constant perfusion biomimetic culture. Lineage specification was assessed by quantitative polymerase chain reaction and immunofluorescent staining. Regenerated left lungs were transplanted in an orthotopic position.
  
  
  RESULTS
  Activin-A treatment, followed by transforming growth factor-β inhibition, induced differentiation of human iPSCs to anterior foregut endoderm as confirmed by forkhead box protein A2 (FOXA2), SRY (Sex Determining Region Y)-Box 17 (SOX17), and SOX2 expression. Cells cultured on decellularized lung slices demonstrated proliferation and lineage commitment after 5 days. Cells expressing Nkx2.1 were identified at 40% to 60% efficiency. Within whole-lung scaffolds and under perfusion culture, cells further upregulated Nkx2.1 expression. After orthotopic transplantation, grafts were perfused and ventilated by host vasculature and airways.
  
  
  CONCLUSIONS
  Decellularized lung matrix supports the culture and lineage commitment of human iPSC-derived lung progenitor cells. Whole-organ scaffolds and biomimetic culture enable coseeding of iPSC-derived endothelial and epithelial progenitors and enhance early lung fate. Orthotopic transplantation may enable further in vivo graft maturation.
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Gilpin
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Xi
authors.initials: X
authors.lastname: Ren
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Okamoto
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Jacques P
authors.initials: JP
authors.lastname: Guyette
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Mou
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Jayaraj
authors.initials: J
authors.lastname: Rajagopal
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Mathisen
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Vacanti
authors.affiliation: Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts. Electronic address: hott@partners.org.
authors.forename: Harald C
authors.initials: HC
authors.lastname: Ott
mesh_terms: D000818:Animals; D021522:Bioartificial Organs; D002102:Cadaver; D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism*; D006085:Graft Survival; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008168:Lung / Q000166:cytology*; D016040:Lung Transplantation / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D054457:Tissue Scaffolds