Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jul 26, 2023; 15(7): 751-767
Published online Jul 26, 2023. doi: 10.4252/wjsc.v15.i7.751
Zinc enhances the cell adhesion, migration, and self-renewal potential of human umbilical cord derived mesenchymal stem cells
Iqra Sahibdad, Shumaila Khalid, G Rasul Chaudhry, Asmat Salim, Sumreen Begum, Irfan Khan
Iqra Sahibdad, Shumaila Khalid, Asmat Salim, Irfan Khan, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Sindh, Pakistan
G Rasul Chaudhry, Department of Biological Sciences, Oakland University, Rochester, MI 48309, United States
Sumreen Begum, Stem Cell Research Laboratory (SCRL), Sindh Institute of Urology and Transplantation (SIUT), Karachi 74200, Sindh, Pakistan
Author contributions: Sahibdad I performed experiments and wrote the original manuscript; Khalid S helped in experimentation and writing; Chaudhry GR evaluated and analyzed the data, and reviewed and edited the manuscript; Salim A evaluated and analyzed the data and reviewed the manuscript; Begum S evaluated and analyzed the data; Khan I conceived and designed the studies, evaluated and analyzed the data, and finalized the manuscript.
Supported by Higher Education Commission, Pakistan indigenous fellowship, No. 520-149004-2MD6-75 (to Ms. Iqra Sahibdad) and No. 520-165117-2BS6-21 (to Ms. Shumaila Khalid).
Institutional review board statement: The independent ethical committee of PCMD, ICCBS, University of Karachi, for human subjects has approved the protocol, No. IEC-009-UCB-2015.
Informed consent statement: Informed consent was obtained from the donor.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data to share.
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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Irfan Khan, PhD, Assistant Professor, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, University Road, Karachi 75270, Sindh, Pakistan. khan@iccs.edu
Received: April 8, 2023
Peer-review started: April 8, 2023
First decision: April 25, 2023
Revised: May 15, 2023
Accepted: June 6, 2023
Article in press: June 6, 2023
Published online: July 26, 2023
Processing time: 107 Days and 21.8 Hours
Abstract
BACKGROUND

Zinc (Zn) is the second most abundant trace element after Fe, present in the human body. It is frequently reported in association with cell growth and proliferation, and its deficiency is considered to be a major disease contributing factor.

AIM

To determine the effect of Zn on in vitro growth and proliferation of human umbilical cord (hUC)-derived mesenchymal stem cells (MSCs).

METHODS

hUC-MSCs were isolated from human umbilical cord tissue and characterized based on immunocytochemistry, immunophenotyping, and tri-lineage differentiation. The impact of Zn on cytotoxicity and proliferation was determined by MTT and Alamar blue assay. To determine the effect of Zn on population doubling time (PDT), hUC-MSCs were cultured in media with and without Zn for several passages. An in vitro scratch assay was performed to analyze the effect of Zn on the wound healing and migration capability of hUC-MSCs. A cell adhesion assay was used to test the surface adhesiveness of hUC-MSCs. Transcriptional analysis of genes involved in the cell cycle, proliferation, migration, and self-renewal of hUC-MSCs was performed by quantitative real-time polymerase chain reaction. The protein expression of Lin28, a pluripotency marker, was analyzed by immunocytochemistry.

RESULTS

Zn at lower concentrations enhanced the rate of proliferation but at higher concentrations (> 100 µM), showed concentration dependent cytotoxicity in hUC-MSCs. hUC-MSCs treated with Zn exhibited a significantly greater healing and migration rate compared to untreated cells. Zn also increased the cell adhesion rate, and colony forming efficiency (CFE). In addition, Zn upregulated the expression of genes involved in the cell cycle (CDC20, CDK1, CCNA2, CDCA2), proliferation (transforming growth factor β1, GDF5, hypoxia-inducible factor 1α), migration (CXCR4, VCAM1, VEGF-A), and self-renewal (OCT4, SOX2, NANOG) of hUC-MSCs. Expression of Lin28 protein was significantly increased in cells treated with Zn.

CONCLUSION

Our findings suggest that zinc enhances the proliferation rate of hUC-MSCs decreasing the PDT, and maintaining the CFE. Zn also enhances the cell adhesion, migration, and self-renewal of hUC-MSCs. These results highlight the essential role of Zn in cell growth and development.

Keywords: Human umbilical cord; Mesenchymal stem cells; Zinc; Cell proliferation; In vitro expansion

Core Tip: Zinc (Zn) efficiently increased the proliferation of mesenchymal stem cells (MSCs) with a significant increase in the rate of population doublings, and at higher concentrations, it was cytotoxic. MSCs of the test group maintained a higher frequency of mesenchymal progenitors. Upregulation of genes involved in cell cycle, proliferation, pluripotency, and migration, was also observed in test groups. Zn increased the migration capability of MSCs and cell adhesion ability of MSCs. Overall, the results showed that Zn at lower concentrations supported cell growth, division, and proliferation of MSCs while maintaining the MSC-specific characteristics, including pluripotency, migration, and cell adhesion.