Original articleGeneral thoracicEnhanced Lung Epithelial Specification of Human Induced Pluripotent Stem Cells on Decellularized Lung Matrix
Section snippets
Human iPSC Culture
All experiments used iPSCs generated by messenger RNA transfection (Kruppel-like factor 4 [Klf4], c-Myc, octamer-binding transcription factor 4 [Oct4], Sox2, LIN28) of postnatal fibroblasts [25]. Undifferentiated cells were maintained in feeder-free culture in mTeSR media (STEMCELL Technologies Inc, Vancouver, BC, Canada) on Geltrex-coated plates (Life Technologies Corp, Carlsbad, CA), and passaged with Accutase (STEMCELL Technologies Inc) at 70% confluency. Differentiation was initiated at 80%
Results
Human fibroblast–derived iPSCs were differentiated in vitro toward a lung epithelial progenitor phenotype through defined developmental stages (Fig 1A). First, activin A treatment generated definitive endoderm at day 4, as confirmed by loss of OCT4 and induction SOX17 and FOXA2 (Fig 1B, C). Next, inhibition of TGF-β signalling for 4 days facilitated anteriorization to a foregut fate, indicated by SOX2 upregulation (Fig 1D, E). Stimulation with FGF-2 and BMP-4, in combination with glycogen
Comment
There is continued need for alternate and novel therapies for patients with end-stage lung disease. Although advances in organ preservation, recipient management, and immunosuppression represent significant therapeutic advancements, lung transplantation remains the only effective treatment option 1, 27. Progress in donor management and organ reconditioning have increased the usable fraction of the donor pool to nearly 50% [28], yet half of all donor lungs are not transplanted, often for
References (37)
- et al.
An overview of tissue and whole organ decellularization processes
Biomaterials
(2011) - et al.
Perfusion decellularization of human and porcine lungs: bringing the matrix to clinical scale
J Heart Lung Transplant
(2014) - et al.
Generation of beta cells from human pluripotent stem cells: Potential for regenerative medicine
Semin Cell Dev Biol
(2012) - et al.
Generation of multipotent lung and airway progenitors from mouse ESCs and patient-specific cystic fibrosis iPSCs
Cell Stem Cell
(2012) - et al.
Defects in tracheoesophageal and lung morphogenesis in Nkx2.1(-/-) mouse embryos
Dev Biol
(1999) - et al.
Preparing for the first breath: genetic and cellular mechanisms in lung development
Dev Cell
(2010) - et al.
Efficient derivation of purified lung and thyroid progenitors from embryonic stem cells
Cell Stem Cell
(2012) - et al.
Repopulation of a human alveolar matrix by adult rat type II pneumocytes in vitro. A novel system for type II pneumocyte culture
Exp Cell Res
(1986) - et al.
Epithelial cells and fibroblasts: structural repair and remodelling in the airways
Paediatr Respir Rev
(2004) - et al.
Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA
Cell Stem Cell
(2010)
Comparison of surgical procedures for vascular and airway anastomoses that utilize a modified non-suture external cuff technique for experimental lung transplantation in rats
J Heart Lung Transplant
The Registry of the International Society for Heart and Lung Transplantation: 29th adult lung and heart-lung transplant report-2012
J Heart Lung Transplant
Vascular endothelial growth factor co-ordinates proper development of lung epithelium and vasculature
Mech Dev
Epithelial-vascular cross talk mediated by VEGF-A and HGF signaling directs primary septae formation during distal lung morphogenesis
Dev Biol
Overview of clinical lung transplantation
Cold Spring Harb Perspect Med
Regeneration and orthotopic transplantation of a bioartificial lung
Nat Med
Enhanced in vivo function of bioartificial lungs in rats
Ann Thorac Surg
Can stem cells be used to generate new lungs? Ex vivo lung bioengineering with decellularized whole lung scaffolds
Respirology
Cited by (109)
Development of lung tissue models and their applications
2023, Life SciencesFuture of Lung Transplantation: Xenotransplantation and Bioengineering Lungs
2023, Clinics in Chest MedicineUnravelling the molecular mechanisms underlying chronic respiratory diseases for the development of novel therapeutics via in vitro experimental models
2022, European Journal of PharmacologyInduced pluripotent stem cells: Novel concepts for respiratory disease modeling
2022, Novel Concepts in iPSC Disease Modeling