Conditioned Media Derived from Human Adipose Tissue Mesenchymal Stromal Cells Improves Primary Hepatocyte Maintenance
Authors: Abbas Piryaei
Hossein Baharvand
Massoud Vosough
Mohamad Reza Hajizadeh
Zahra Azhdari Tafti
Vahid Ezzatizadeh
Mehdi Mahmoodi
Journal: Cell journal (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Conditioned Medium; Mesenchymal Stromal Cell; Primary Hepatocyte; Regenerative Medicine.
Affiliations: Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran., Department of Clinical Biochemistry, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran., Molecular Medicine Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran., Department of Medical Genetics, Medical Laboratory Center, Royesh Medical Group, Tehran, Iran., Department of Developmental Biology, University of Science and Culture, Tehran, Iran., Department of Regenerative Biomedicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Electronic Address:., Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic Address:.

Abstract:  Objective: Recent advances in cell therapy have encouraged researchers to provide an alternative for treatment and restoration of damaged liver through using hepatocytes. However, these cells quickly lose their functional capabilities in vitro. Here, we aim to use the secretome of mesenchymal stromal cells (MSCs) to improve in vitro maintenance conditions for hepatocytes. Materials and methods: In this experimental study, following serum deprivation, human adipose tissue-derived MSCs (hAT-MSCs) were cultured for 24 hours under normoxic (N) and hypoxic (H) conditions. Their conditioned media (CM) were subsequently collected and labeled as N-CM (normoxia) and H-CM (hypoxia). Murine hepatocytes were isolated by perfusion of mouse liver with collagenase, and were cultured in hepatocyte basal (William's) medium supplemented with 4% N-CM or H-CM. Untreated William's and hepatocyte-specific media (HepZYM) were used as controls. Finally, we evaluated the survival and proliferation rates, as well as functionality and hepatocyte-specific gene expressions of the cells. Results: We observed a significant increase in viability of hepatocytes in the presence of N-CM and H-CM compared to HepZYM on day 5, as indicated by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium) assay. Indocyanine green (ICG) uptake of hepatocytes in the H-CM and HepZYM groups on days 3 and 5 also suggested that H-CM maintained the hepatocytes at about the same level as the hepatocyte-specific medium. The HepZYM group had significantly higher levels of albumin (Alb) and urea secretion compared to the other groups (P&lt;0.0001). However, there were no significant differences in cytochrome activity and cytochrome gene expression profiles among these groups. Finally, we found a slightly, but not significantly higher concentration of vascular endothelial growth factor (VEGF) in the H-CM group compared to the N-CM group (P=0.063). Conclusion: The enrichment of William's basal medium with 4% hAT-MSC-H-CM improved some physiologic parameters in a primary hepatocyte culture. 

Retracted: No

PubPeer comments:
 Figure 5A. * Some panels appear to show a small duplicated area in the bottom right corner. Indicated here with boxes of the same color. Could the authors please clarify what happened here? ![file] , 
 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. We agree with your concern. However, we checked the proofed version before production. It was correct. Then we contacted the journal and they replied the following and also the journal was sent its email to you before. The journal informed us: u201cAfter assessing our document, we found out that in the original panel received from the authors, three figures had scale bars, and the journal s graphic designer for uniformity of the figures, removed the scale bars from the bottom right corner of these three figures and copy-pasted from the neighboring sides to restore the blank part. She did it without the permission of the authors and as it was done at the last step after the final checking of the authors, so this mistake takes place in your paper. We apologize for this mistake, and any inconvenience caused. We will publish a correction in the next issue to inform the readers.u201d , 
 #3 u201c[T]he journal s graphic designer for uniformity of the figures, removed the scale bars from the bottom right corner of these three figures and copy-pasted from the neighboring sides to restore the blank part.u201d Astonishing. I would note that the responding author appears to be on the editorial board of this journal. https: celljournal.org about-journal , 
 #4 Really sorry instead of paying attention to the cell journal answer, you are accusing the journal to be not honest, because Prof. Baharvand is one of the editorial members of the journal. Would you please describe what is the benefit of authors to duplicate a tiny slice of the image from the middle bottom to the right bottom of the image? H. Gourabi Chairman of Cell Journal , 
 #5 I apologize for confusing you. I did not describe anyoneu2019s actions as dishonest and the answer does satisfactorily explain how and why this occurred. So thank you for that. I sincerely have never heard of journal staff altering original data without permission, or without the cooperation, of the authors, and was merely expressing my surprise. , 
 #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: www.ncbi.nlm.nih.gov pmc articles PMC7944132  In this article which was published in Cell J, Vol 20, No 3, Autumn 2018, on pages 377-387, the scale bars in Figures 5-A missed unintentionally during production. The following figure is corrected. The authors would like to apologies for any inconvenience caused. It is still not clear to me what has happened. This is not a simple case of missing scale bars, but the correction does not mention the cloned photo features. ,