Regenerative potential of human airway stem cells in lung epithelial engineering.
Bio-engineered organs for transplantation may ultimately provide a personalized solution for end-stage organ failure, without the risk of rejection. Building upon the process of whole organ perfusion decellularization, we aimed to develop novel, translational methods for the recellularization and regeneration of transplantable lung constructs. We first isolated a proliferative KRT5(+)TP63(+) basal epithelial stem cell population from human lung tissue and demonstrated expansion capacity in conventional 2D culture. We then repopulated acellular rat scaffolds in ex vivo whole organ culture and observed continued cell proliferation, in combination with primary pulmonary endothelial cells. To show clinical scalability, and to test the regenerative capacity of the basal cell population in a human context, we then recellularized and cultured isolated human lung scaffolds under biomimetic conditions. Analysis of the regenerated tissue constructs confirmed cell viability and sustained metabolic activity over 7 days of culture. Tissue analysis revealed extensive recellularization with organized tissue architecture and morphology, and preserved basal epithelial cell phenotype. The recellularized lung constructs displayed dynamic compliance and rudimentary gas exchange capacity. Our results underline the regenerative potential of patient-derived human airway stem cells in lung tissue engineering. We anticipate these advances to have clinically relevant implications for whole lung bioengineering and ex vivo organ repair.
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Sarah E
SE
Gilpin
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Jonathan M
JM
Charest
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Xi
X
Ren
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Luis F
LF
Tapias
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Tong
T
Wu
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Daniele
D
Evangelista-Leite
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States.
Douglas J
DJ
Mathisen
Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, United States; Harvard Medical School, United States; Center for Regenerative Medicine, Massachusetts General Hospital, United States.
Harald C
HC
Ott
D021522:Bioartificial Organs; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000254:growth & development*; D012038:Regeneration / Q000502:physiology; D020545:Respiratory Mucosa / Q000166:cytology* / Q000502:physiology; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds
Decellularization; Epithelial repair; Extracellular matrix; Lung tissue engineering; Recellularization; Regeneration