Author/Authors :
Noori ، Afshin Department of Internal Medicine - Faculty of Veterinary Medicine - University of Tehran , Mokhber Dezfouli ، Mohammad Reza Department of Internal Medicine - Faculty of Veterinary Medicine, Institute of Biomedical Research - University of Tehran , Rajabi ، Sareh Department of Stem Cells and Developmental Biology, Department of Cell Engineering - Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology - Academic Center for Education, Culture and Research (ACECR) , Ganji ، Fatemeh Department of Tissue Engineering and Regenerative Medicine - Faculty of Advanced Technologies in Medicine - Iran University of Medical Sciences , Ghezelayagh ، Zahra Department of Stem Cells and Developmental Biology - Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology - Academic Center for Education, Culture and Research (ACECR) , El Agha ، Elie Department of Internal Medicine - Universities of Giessen and Marburg Lung Center (UGMLC), Institute for Lung Health (ILH), Cardio-Pulmonary Institute (CPI), German Center for Lung (DZL) - Justus-Liebig University Giessen , Baharvand ، Hossein Department of Stem Cells and Developmental Biology - Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology - Academic Center for Education, Culture and Research (ACECR) , Sadeghian Chaleshtori ، Sirous Department of Internal Medicine - Faculty of Veterinary Medicine, Institute of Biomedical Research - University of Tehran , Tahamtani ، Yaser Department of Stem Cells and Developmental Biology - Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine - Academic Center for Education, Culture and Research (ACECR)
Abstract :
Objective: Efficient production of functional and mature alveolar epithelial is a major challenge for developing any cellreplacement therapy for lung degenerative diseases. The extracellular matrix (ECM) pro-vides a dynamic environmentand mediates cellular responses during development and maintenance of tissue functions. The decellularized ECM(dECM) which retains its native-like structure and bio-chemical composition can provide the induction of embryonicstem cell (ESC) differentiation toward the tissue-specific lineages during in vitro culture. Therefore, the aim of this studywas to evaluate the effect of sheep lung dECM-derived scaffold on differentiation and further maturation of ESC-derivedlung progenitor cells.Materials and Methods: This study was an experimental study. In the first step, a sheep lung was decellularizedto achieve dECM scaffolds and hydrogels. Afterwards, the obtained dECM scaffold was evaluated for collagen andglycosaminoglycan contents, DNA quantification, and its ultrastructure. Next, the three experimental groups: i. Sheeplung dECM-derived scaffold, ii. Sheep lung dECM-derived hydrogel, and iii. Fibronectin-coated plates were comparedin their abilities to induce further differentiation of human embryonic stem cells (hESCs)-derived definitive endoderm(DE) into lung progenitor cells. The comparison was evaluated by immuno-staining and real-time polymerase chainreaction (PCR) assessments.Results: We found that the dECM-derived scaffold preserved its composition and native porous structures whilelacking nuclei and intact cells. All experimental groups displayed lung progenitor cell differen-tiation as revealed by theRNA and protein expression of NKX2.1, P63 and CK5. DE cells differenti-ated on dECM-derived scaffold and dECMderivedhydrogel showed significant upregulation of SOX9 gene expression, a marker of the distal airway epithelium.DE cells differentiated on the dECM-derived scaffold compared to the two other groups, showed enhanced expressionof SFTPC (type 2 alveolar epithelial [AT2] cell marker), FOXJ1 (ciliated cell marker), and MUC5A (secretory cell marker)genes.Conclusion: Overall, our results suggest that dECM-derived scaffold improves the differentiation of DE cells towardslung alveolar progenitor cells in comparison with dECM-derived hydrogel and fibronectin-coated plates.
Keywords :
Decellularization , Differentiation , Hydrogel , Lung , Scaffold
