title: Fibroblast engraftment in the decellularized mouse lung occurs via a β1-integrin-dependent, FAK-dependent pathway that is mediated by ERK and opposed by AKT.
abstract: Creation of bioartificial organs has been enhanced by the development of strategies involving decellularized mammalian lung. Because fibroblasts critically support lung function through a number of mechanisms, study of these cells in the context of the decellularized lung has the potential to improve the structure and function of tissue-engineered lungs. We characterized the engraftment and survival of a mouse fibroblast cell line in decellularized rat lung slices and found a time-dependent increase in cell numbers assessed by hematoxylin and eosin staining, cell proliferation assessed by Ki67 staining, and minimal cell death assessed by TUNEL staining. We developed a repopulation index to allow quantification of cell survival that accounts for variation in cell density throughout the seeded scaffold. We then applied this method to the study of mouse lung scaffolds and found that decellularization of presliced mouse lungs produced matrices with preserved alveolar architecture and proteinaceous components including fibronectin, collagens I and IV, laminin, and elastin. Treatment with a β1-integrin-neutralizing antibody significantly reduced the repopulation index after 24 h of culture. Treatment with focal adhesion kinase (FAK) inhibitor and extracellular signal-regulated kinase (ERK) inhibitor further reduced initial repopulation scores while treatment with AKT inhibitor increased initial scores. Rho-associated kinase inhibitor had no discernible effect. These data indicate that initial adhesion and survival of mouse fibroblasts in the decellularized mouse lung occur in a β1-integrin-dependent, FAK/ERK-dependent manner that is opposed by AKT.
authors: Yale School of Medicine, Section of Pulmonary, Critical Care, and Sleep Medicine, 300 Cedar St. TAC 441S, New Haven CT 06520-8057. erica.herzog@yale.edu.
authors: Huanxing
authors: H
authors: Sun
authors: Elizabeth
authors: E
authors: Calle
authors: Xiaosong
authors: X
authors: Chen
authors: Aditi
authors: A
authors: Mathur
authors: Yangyang
authors: Y
authors: Zhu
authors: Julio
authors: J
authors: Mendez
authors: Liping
authors: L
authors: Zhao
authors: Laura
authors: L
authors: Niklason
authors: Xueyan
authors: X
authors: Peng
authors: Hong
authors: H
authors: Peng
authors: Erica L
authors: EL
authors: Herzog
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D021522:Bioartificial Organs; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005347:Fibroblasts / Q000502:physiology / Q000637:transplantation*; D051417:Focal Adhesion Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019012:Integrin beta1 / Q000276:immunology / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000502:physiology*; D051379:Mice; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors
chemical_list: D057134:Antibodies, Neutralizing; D019012:Integrin beta1; D051417:Focal Adhesion Kinase 1; C496051:Ptk2 protein, mouse; D051057:Proto-Oncogene Proteins c-akt; D054460:rho-Associated Kinases; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: bioengineering; decellularized lung; fibroblast