Hydrocortisone Promotes Differentiation of Mouse Embryonic Stem Cell-Derived Definitive Endoderm toward Lung Alveolar Epithelial Cells
Authors: Hossein Baharvand
Azadeh Moradmand
Mohammad Reza Mokhber Dezfouli
Sirous Sadeghian Chaleshtori
Mohsen Basiri
Yaser Tahamtani
Journal: Cell journal (2019)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Differentiation; Embryonic Stem Cells; Lung; Regenerative Medicine.
Affiliations: Department of Internal Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran., Institute of Biomedical Research, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. Electronic Address: ., Institute of Biomedical Research, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran., Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran., Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran., Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell. Electronic Address: .

Abstract:  Objective: The ability to generate lung alveolar epithelial type II (ATII) cells from pluripotent stem cells (PSCs) enables the study of lung development, regenerative medicine, and modeling of lung diseases. The establishment of defined, scalable differentiation methods is a step toward this goal. This study intends to investigate the competency of small molecule induced mouse embryonic stem cell-derived definitive endoderm (mESC-DE) cells towards ATII cells. Materials and methods: In this experimental study, we designed a two-step differentiation protocol. mESC line Royan B20 (RB20) was induced to differentiate into DE (6 days) and then into ATII cells (9 days) by using an adherent culture method. To induce differentiation, we treated the mESCs for 6 days in serum-free differentiation (SFD) media and induced them with 200 nM small molecule inducer of definitive endoderm 2 (IDE2). For days 7-15 (9 days) of induction, we treated the resultant DE cells with new differentiation media comprised of 100 ng/ml fibroblast growth factor (FGF2) (group F), 0.5 Œºg/ml hydrocortisone (group H), and A549 conditioned medium (A549 CM) (group CM) in SFD media. Seven different combinations of factors were tested to assess the efficiencies of these factors to promote differentiation. The expressions of DE- and ATII-specific markers were investigated during each differentiation step. Results: Although both F and H (alone and in combination) promoted differentiation through ATII-like cells, the highest percentage of surfactant protein C (SP-C) expressing cells (~37%) were produced in DE-like cells treated by F+H+CM. Ultrastructural analyses also confirmed the presence of lamellar bodies (LB) in the ATII-like cells. Conclusion: These results suggest that hydrocortisone can be a promoting factor in alveolar fate differentiation of IDE2-induced mESC-DE cells. These cells have potential for drug screening and cell-replacement therapies. 

Retracted: No

PubPeer comments:
 The Acknowledgments section of this paper includes the statement:  The authors have no conflicts of interest with the content of this article. Yet, this paper appears to show a significant overlap with patent application US20180051257A1, which was filed on 18 July 2017 according to https: patents.google.com patent US20180051257A1 , while this paper was submitted on 25 September 2017, almost a month later. Abstract, text, and figures appear to look very similar. Here, for example, is Figure 1 from this paper (left) compared to Figures 4A-7B from the patent application.  Could the authors please clarify why they think there was no conflict of interest to declare here? , 
 Dear Dr. Bik, Thank you so much for your analysis of my articles! Currently, we are carefully checking each of your comments and concerns regarding  image concerns and undisclosed conflict of interest in some of my papers. We have also started to contact the related journals to ask for corrective notes on those articles that need correction or other potential actions. We will update, on Pubpeer, the final changes and actions communicated with the journals once we get feedback from the journals. Thank you again for your time and concern! Sincerely yours, Hossein Baharvand , 
 #1 Thank you for your concern. Correspondents have explained to the editor and asked to publish an Erratum to our manuscript to avoid any potential ambiguities in the future and they accepted. , 
 Correction, April 2021: https: pubmed.ncbi.nlm.nih.gov 33650830  In this article which was published in Cell J, Vol 20, No 4, winter 2019, on pages 469-479, the authors regret to acknowledge that we failed to mention in our article that a patent based on this study had been filed by Royan Institute and Tehran University with S.S.C., M.R.M.D., H.B., and Y.T. as inventors. The authors would like to apologies for any inconvenience caused. ,