Enhanced reseeding of decellularized rodent lungs with mouse embryonic stem cells.
Repopulation of decellularized lung scaffolds (DLS) is limited due to alterations in the repertoire and ratios of the residual extracellular matrix (ECM) proteins, characterized by e.g., the retention of type I collagen and loss of glycoproteins. We hypothesized that pre-treatment of decellularized matrices with defined ECM proteins, which match the repertoire of integrin receptors expressed by the cells to be seeded (e.g., embryonic stem cells) can increase the efficacy of the reseeding process. To test this hypothesis, we first determined the integrin receptors profile of mouse embryonic stem cells (mESCs). Mouse ESCs express α3, α5, α6, α9 and β1, but not α1, α2 and α4 integrin subunits, as established by Western blotting and adhesion to laminin and fibronectin, but not to collagens type I and IV. Reseeding of DLS with mESCs was inefficient (6.9 ± 0.5%), but was significantly enhanced (2.3 ± 0.1 fold) by pre-treating the scaffolds with media conditioned by A549 human lung adenocarcinoma cells, which we found to contain ∼5 μg/ml laminin. Furthermore, pre-treatment with A549-conditioned media resulted in a significantly more uniform distribution of the seeded mESCs throughout the engineered organ as compared to untreated DLS. Our study may advance whole lung engineering by stressing the importance of matching the integrin receptor repertoire of the seeded cells and the cell binding motifs of DLS.
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA.
Shimon
S
Lecht
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA.
Collin T
CT
Stabler
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA.
Alexis L
AL
Rylander
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA.
Rachel
R
Chiaverelli
Division of Pulmonary, Critical Care and Sleep Medicine, Drexel University College of Medicine, Philadelphia, PA 19102, USA.
Edward S
ES
Schulman
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA.
Cezary
C
Marcinkiewicz
Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA 19122, USA. Electronic address: pilelkes@temple.edu.
Peter I
PI
Lelkes
D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion; D002460:Cell Line; D017077:Culture Media, Conditioned; D053595:Embryonic Stem Cells / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000166:cytology*; D051379:Mice; D033581:Stem Cell Transplantation
D017077:Culture Media, Conditioned; D016326:Extracellular Matrix Proteins
Acellular matrix; Cell adhesion; Embryonic stem cells; Extracellular matrix (ECM); Integrin; Lung