Immortalization of epidural fat-derived mesenchymal stem cells: In vitro characterization and adipocyte differentiation potential

BACKGROUND

Mesenchymal stem cells (MSCs) are promising candidates for regenerative therapy due to their self-renewal capability, multilineage differentiation potential, and immunomodulatory effects. The molecular characteristics of MSCs are influenced by their location. Recently, epidural fat (EF) and EF-derived MSCs (EF-MSCs) have garnered attention due to their potential benefits to the spinal microenvironment and their high expression of neural SC markers. However, their clinical applications are limited due to cell senescence and limited accessibility of EF. Although many studies have attempted to establish an immortalized, stable SC line, the characteristics of immortalized EF-MSCs remain to be clarified.

AIM

To establish and analyze stable immortalized EF-MSCs.

METHODS

The phenotypes of EF-MSCs were analyzed using optical microscopy. Cell immortalization was performed using lentiviral vectors. The biomolecular characteristics of the cells were analyzed by immunoblotting, quantitative PCR, and proteomics.

RESULTS

The immortalized EF-MSCs demonstrated a significantly extended lifespan compared to the control group, with well-preserved adipogenic potential and SC surface marker expression. Introduction of human telomerase reverse transcriptase genes markedly increased the lifespan of EF-MSCs. Proteomics analysis revealed substantial increase in the expression of DNA replication pathway components in immortalized EF-MSCs.

CONCLUSION

Immortalized EF-MSCs exhibited significantly enhanced proliferative capacity, retained adipogenic potential, and upregulated the expression of DNA replication pathway components.

Key Words: Immortalization; Transfection; Stem cell; Epidural fat; Proteomics

Core Tip: Epidural fat-derived mesenchymal stem cells (EF-MSCs) are potential candidates to treat neurodegenerative diseases. However, the clinical implication of EF-MSCs has encountered many limitations including difficulty in isolating the fat from the epidural region and limited proliferation ability of the SCs. Therefore, precise biomolecular characteristics of the EF-MSC remain unclear. To overcome these problems, we immortalized EF-MSCs through lentiviral transduction, and these stable EF-MSCs were analyzed by phenotypical interpretation and proteomics. In this study, immortalized EF-MSCs demonstrated notably increased lifespan and adipogenic potential. These findings suggest that stable EF-MSCs could be potential tools for developing effective regenerative medicine strategies.

• Citation: Lee SW, Lim YJ, Kim HY, Kim W, Park WT, Ma MJ, Lee J, Seo MS, Kim YI, Park S, Choi SK, Lee GW. Immortalization of epidural fat-derived mesenchymal stem cells: In vitro characterization and adipocyte differentiation potential. World J Stem Cells 2025; 17(1): 98777
• URL: https://www.wjgnet.com/1948-0210/full/v17/i1/98777.htm
• DOI: https://dx.doi.org/10.4252/wjsc.v17.i1.98777