Basic Study
Copyright ©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jan 26, 2025; 17(1): 101485
Published online Jan 26, 2025. doi: 10.4252/wjsc.v17.i1.101485
Extended protective effects of three dimensional cultured human mesenchymal stromal cells in a neuroinflammation model
Ok-Hyeon Kim, Hana Kang, Eun Seo Chang, Younghyun Lim, Young-Jin Seo, Hyun Jung Lee
Ok-Hyeon Kim, Hyun Jung Lee, Department of Anatomy and Cell Biology, College of Medicine, Chung-Ang University, Seoul 06974, South Korea
Hana Kang, Eun Seo Chang, Hyun Jung Lee, Department of Global Innovative Drugs, Graduate School of Chung-Ang University, Seoul 06974, South Korea
Younghyun Lim, Young-Jin Seo, Department of Life Science, Chung-Ang University, Seoul 06974, South Korea
Author contributions: Kim OH, Kang H, Chang ES, Lim Y, Seo YJ, and Lee H contributed to the methodology; Kim OH and Lee HJ contributed to the conceptualization and writing-original draft; Kim OH, Kang H, Chang ES, and Lim Y contributed to the formal analysis; Kim OH contributed to validation and investigation; Seo YJ and Lee HJ contributed to supervision; Lee HJ contributed to review & editing, project administration, and funding acquisition.
Supported by National Research Foundation of Korea, No. RS-2024-00409554, No. 2023R1A2C2006894, and No. 2021R1A6A3A01088243.
Institutional animal care and use committee statement: All experimental procedures including animals were reviewed and approved by the Institutional Animal Care Use Committee of Chung-Ang University (Approval ID: A2021023) and performed in accordance with the Guide for the Care and Use of Laboratory Animals, 8th edition (NIH publication No. 85-23, National Academy Press, Washington, DC, United States, revised 2011).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: All data generated or analyzed during this study are included in this published article and its supplementary information files.
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: Hyun Jung Lee, PhD, Department of Anatomy and Cell Biology, College of Medicine, Chung-Ang University, 84 Heuksuk-ro, Dongjak-gu, Seoul 06974, South Korea. pluto38@cau.ac.kr
Received: September 16, 2024
Revised: November 11, 2024
Accepted: January 8, 2025
Published online: January 26, 2025
Processing time: 125 Days and 22 Hours
Abstract
BACKGROUND

Human mesenchymal stromal cells (MSCs) possess regenerative potential due to pluripotency and paracrine functions. However, their stemness and immunomodulatory capabilities are sub-optimal in conventional two-dimensional (2D) culture.

AIM

To enhance the efficiency and therapeutic efficacy of MSCs, an in vivo-like 3D culture condition was applied.

METHODS

MSCs were cultured on polystyrene (2D) or in a gellan gum-based 3D system. In vitro, prostaglandin-endoperoxide synthase 2, indoleamine-2,3-dioxygenase, heme oxygenase 1, and prostaglandin E synthase gene expression was quantified by quantitative real-time polymerase chain reaction. MSCs were incubated with lipopolysaccharide (LPS)-treated mouse splenocytes, and prostaglandin E2 and tumor necrosis factor-alpha levels were measured by enzyme linked immunosorbent assay. In vivo, LPS was injected into the lateral ventricle of mouse brain, and MSCs were administered intravenously the next day. Animals were sacrificed and analyzed on days 2 and 6.

RESULTS

Gellan gum polymer-based 3D culture significantly increased expression of octamer-binding transcription factor 4 and Nanog homeobox stemness markers in human MSCs compared to 2D culture. This 3D environment also heightened expression of cyclooxygenase-2 and heme-oxygenase 1, enzymes known for immunomodulatory functions, including production of prostaglandins and heme degradation, respectively. MSCs in 3D culture secreted more prostaglandin E2 and effectively suppressed tumor necrosis factor-alpha release from LPS-stimulated splenocytes and surpassed the efficiency of MSCs cultured in 2D. In a murine neuroinflammation model, intravenous injection of 3D-cultured MSCs significantly reduced ionized calcium-binding adaptor molecule 1 and glial fibrillary acidic protein expression, mitigating chronic inflammation more effectively than 2D-cultured MSCs.

CONCLUSION

The microenvironment established in 3D culture serves as an in vivo mimetic, enhancing the immunomodulatory function of MSCs. This suggests that engineered MSCs hold significant promise a potent tool for cell therapy.

Keywords: Mesenchymal stromal cells; Three-dimensional culture; Immunomodulatory function; Neuroinflammation; Cell therapy

Core Tip: Clinical trials utilizing mesenchymal stromal cells (MSCs) have been on the rise, but several barriers to broader adoption of MSCs as therapeutics remain to be addressed. Our study presents compelling evidence demonstrating the remarkable enhancement of immunomodulatory capabilities in functional polymer-based MSCs through three-dimensional culture. We specifically applied these optimized MSCs to a brain inflammation model and found a prolonged and more potent anti-inflammatory effect in vivo compared to conventionally cultured MSCs. The polymer-based three-dimensional cultures we propose could be an easy tool for mass culture for clinical applications.