Fatemeh Ganji
12 
, Marzieh Ebrahimi
3, Ali Shirani
1,2, Mahtab Golmohammadi
4, Mazaher Gholipourmalekabadi
1,2, Maryam Kashanian
5, Kiana Koolaeinezhad
5, Hamid Reza Davari
6, Seyed Ali Javad Mousavi
7, Hamid Reza Aghayan
8, Babak Arjmand
8,9, Ramin Heshmat
10, Nushin Karkuki Osguei
11, Ali Samadikuchaksaraei
12* 1 Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
2 Department of Tissue Engineering & Regenerative Medicine, Iran University of Medical Sciences, Tehran, Iran
3 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
4 Department of Genetic, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
5 Shahid-Akbarabadi Clinical Research Development Unit, Iran University of Medical Sciences, Tehran, Iran
6 Thorax Advanced Research Center, Tehran University of Medical Sciences, Tehran, Iran
7 Section of Respiratory Diseases, Department of Internal Medicine, Iran University of Medical Sciences, Tehran, Iran
8 Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular-Cellular. Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran
9 Metabolomics and Genomics Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran
10 Chronic Diseases Research Center, Tehran University of Medical Sciences, Tehran, Iran
11 Eposcience Millennium Institute, Tehran, Iran
12 Department of Medical Biotechnology, Iran University of Medical Sciences, Tehran, Iran
Abstract
Introduction: For cell-based therapies of lung injury, several cell sources have been extensively studied. However, the potential of human fetal respiratory cells has not been systematically explored for this purpose. Here, we hypothesize that these cells could be one of the top sources and hence, we extensively updated the definition of their phenotype.
Methods: Human fetal lower respiratory tissues from pseudoglandular and canalicular stages and their isolated epithelial cells were evaluated by immunostaining, electron microscopy, flow cytometry, organoid assay, and gene expression studies. The regenerative potential of the isolated cells has been evaluated in a rat model of bleomycin-induced pulmonary injury by tracheal instillation on days 0 and 14 after injury and harvest of the lungs on day 28.
Results: We determined the relative and temporal, and spatial pattern of expression of markers of basal (KRT5, KRT14, TRP63), non-basal (AQP3 and pro-SFTPC), and early progenitor (NKX2.1, SOX2, SOX9) cells. Also, we showed the potential of respiratory-derived cells to contribute to in vitro formation of alveolar and airway-like structures in organoids. Cell therapy decreased fibrosis formation in rat lungs and improved the alveolar structures. It also upregulated the expression of IL-10 (up to 17.22 folds) and surfactant protein C (up to 2.71 folds) and downregulated the expression of TGF-β (up to 5.89 folds) and AQP5 (up to 3.28 folds).
Conclusion: We provide substantial evidence that human fetal respiratory tract cells can improve the regenerative process after lung injury. Also, our extensive characterization provides an updated phenotypic profile of these cells.