Extended Protective Effects of Three Dimensional Cultured Human Mesenchymal Stromal Cells in a Neuroinflammation Model

Overview

Journal World J Stem Cells

Publisher Baishideng Publishing Group

Date 2025 Jan 27

PMID 39866897

Authors

Ok-Hyeon Kim

Hana Kang

Eun Seo Chang

Younghyun Lim

Young-Jin Seo

Hyun Jung Lee

Affiliations

Soon will be listed here.

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 -like 3D culture condition was applied.

Methods: MSCs were cultured on polystyrene (2D) or in a gellan gum-based 3D system. , 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. , 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 mimetic, enhancing the immunomodulatory function of MSCs. This suggests that engineered MSCs hold significant promise a potent tool for cell therapy.

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