Decellularized Lung Extracellular Matrix Scaffold Promotes Human Embryonic Stem Cell Differentiation towards Alveolar Progenitors

Document Type : Original Article


1 Department of Internal Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

3 Institute of Biomedical Research, University of Tehran, Tehran, Iran

4 Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

5 Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran

6 Faculty of Sciences and Advanced Technologies in Biology, University of Science and Culture, ACECR, Tehran, Iran

7 Department of Internal Medicine, Universities of Giessen and Marburg Lung Center (UGMLC), Institute for Lung Health (ILH), Cardio-Pulmonary Institute (CPI), Member of the German Center for Lung (DZL), Justus-Liebig University Giessen, Giessen, Germany

8 Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran


Objective: Efficient production of functional and mature alveolar epithelial is a major challenge for developing any cell
replacement therapy for lung degenerative diseases. The extracellular matrix (ECM) pro-vides a dynamic environment
and 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 embryonic
stem cell (ESC) differentiation toward the tissue-specific lineages during in vitro culture. Therefore, the aim of this study
was to evaluate the effect of sheep lung dECM-derived scaffold on differentiation and further maturation of ESC-derived
lung progenitor cells.
Materials and Methods: This study was an experimental study. In the first step, a sheep lung was decellularized
to achieve dECM scaffolds and hydrogels. Afterwards, the obtained dECM scaffold was evaluated for collagen and
glycosaminoglycan contents, DNA quantification, and its ultrastructure. Next, the three experimental groups: i. Sheep
lung dECM-derived scaffold, ii. Sheep lung dECM-derived hydrogel, and iii. Fibronectin-coated plates were compared
in 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 chain
reaction (PCR) assessments.
Results: We found that the dECM-derived scaffold preserved its composition and native porous structures while
lacking nuclei and intact cells. All experimental groups displayed lung progenitor cell differen-tiation as revealed by the
RNA and protein expression of NKX2.1, P63 and CK5. DE cells differenti-ated on dECM-derived scaffold and dECMderived
hydrogel 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 expression
of SFTPC (type 2 alveolar epithelial [AT2] cell marker), FOXJ1 (ciliated cell marker), and MUC5A (secretory cell marker)
Conclusion: Overall, our results suggest that dECM-derived scaffold improves the differentiation of DE cells towards
lung alveolar progenitor cells in comparison with dECM-derived hydrogel and fibronectin-coated plates.


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