Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Oct 26, 2021; 13(10): 1580-1594
Published online Oct 26, 2021. doi: 10.4252/wjsc.v13.i10.1580
Effect of glycyrrhizic acid and 18β-glycyrrhetinic acid on the differentiation of human umbilical cord-mesenchymal stem cells into hepatocytes
Abiha Fatima, Tuba Shakil Malick, Irfan Khan, Aisha Ishaque, Asmat Salim
Abiha Fatima, Tuba Shakil Malick, Irfan Khan, Aisha Ishaque, Asmat Salim, Stem Cell Laboratory, Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Sindh, Pakistan
Author contributions: Fatima A performed all experiments, analyzed the data and wrote the first draft; Malick TS and Ishaque A performed part of the experiments and analysis of the data; Khan I provided technical guidance for all experiments and data analysis; Salim A designed the research study, supervised the research and wrote the manuscript in its final form; all authors have read and approved the final manuscript.
Institutional review board statement: Present study was approved by the Independent Ethics Committee, International Center for Chemical and Biological Sciences (IEC, ICCBS) under protocol #: ICCBS/IEC-037-HT-2018/Protocol/1.0.
Conflict-of-interest statement: The authors confirm that this article has no conflict of interest to report.
Data sharing statement: No additional data is available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Asmat Salim, PhD, Professor, Stem Cell Laboratory, Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Sindh, Pakistan. asmat.salim@iccs.edu
Received: April 13, 2021
Peer-review started: April 13, 2021
First decision: May 12, 2021
Revised: May 25, 2021
Accepted: September 19, 2021
Article in press: September 19, 2021
Published online: October 26, 2021
Processing time: 195 Days and 17.4 Hours
Abstract
BACKGROUND

End-stage liver disease is a global health complication with high prevalence and limited treatment options. Cell-based therapies using mesenchymal stem cells (MSCs) emerged as an alternative approach to support hepatic regeneration. In vitro preconditioning strategies have been employed to strengthen the regenerative and differentiation potential of MSCs towards hepatic lineage. Chemical compounds of the triterpene class; glycyrrhizic acid (GA) and 18β-glycyrrhetinic acid (GT) possess diverse therapeutic properties including hepato-protection and anti-fibrosis characteristics. They are capable of modulating several signaling pathways that are crucial in hepatic regeneration. Preconditioning with hepato-protective triterpenes may stimulate MSC fate transition towards hepatocytes.

AIM

To explore the effect of GA and GT on hepatic differentiation of human umbilical cord-MSCs (hUC-MSCs).

METHODS

hUC-MSCs were isolated and characterized phenotypically by flow cytometry and immunocytochemistry for the expression of MSC-associated surface molecules. Isolated cells were treated with GA, GT, and their combination for 24 h and then analyzed at three time points; day 7, 14, and 21. qRT-PCR was performed for the expression of hepatic genes. Expression of hepatic proteins was analyzed by immunocytochemistry at day 21. Periodic acid Schiff staining was performed to determine the functional ability of treated cells.

RESULTS

The fusiform-shaped morphology of MSCs in the treatment groups in comparison with the untreated control, eventually progressed towards the polygonal morphology of hepatocytes with the passage of time. The temporal transcriptional profile of preconditioned MSCs displayed significant expression of hepatic genes with increasing time of differentiation. Preconditioned cells showed positive expression of hepatocyte-specific proteins. The results were further corroborated by positive periodic acid Schiff staining, indicating the presence of glycogen in their cytoplasm. Moreover, bi-nucleated cells, which is the typical feature of hepatocytes, were also seen in the preconditioned cells.

CONCLUSION

Preconditioning with glycyrrhizic acid, 18β-glycyrrhetinic acid and their combination, successfully differentiates hUC-MSCs into hepatic-like cells. These MSCs may serve as a better therapeutic option for degenerative liver diseases in future.

Keywords: Glycyrrhizic acid, 18β-glycyrrhetinic acid, Hepatocyte differentiation, Human umbilical cord-MSCs, Mesenchymal stem cells

Core Tip: This study focuses on exploring the effect of two triterpenes, glycyrrhizic acid and 18β-glycyrrhetinic acid to enhance the differentiation potential of mesenchymal stem cells (MSCs) into hepatocytes to ensure a potent and valuable cell source for cellular therapy for end-stage liver disease. Preconditioning of human umbilical cord-MSCs with these compounds enhances expression of both early and late hepatic markers, regulated with time of differentiation. The significant expression of hepatocyte markers, the ability to store glycogen, and presence of bi-nucleated cells, suggest successful hepatic differentiation. Preconditioned MSCs may help in the replacement of damaged hepatocytes, and improve liver function post-transplantation in impaired hepatic tissues.