Lee Seoung-Woo, Lim Young-Ju, Kim Hee-Yeon, Kim Wansoo, Park Wook-Tae, Ma Min-Jung, Lee Junho, Seo Min-Soo, Kim Young In, Park Sangbum, Choi Seong-Kyoon, Lee Gun Woo
Core Protein Resources Center, Daegu-Gyeongbuk Institute of Science and Technology, Daegu 42988, South Korea.
Department of Orthopedic Surgery, Yeungnam University Medical Center, Yeungnam University College of Medicine, Daegu 42415, South Korea.
World J Stem Cells. 2025 Jan 26;17(1):98777. doi: 10.4252/wjsc.v17.i1.98777.
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.
To establish and analyze stable immortalized EF-MSCs.
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.
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.
Immortalized EF-MSCs exhibited significantly enhanced proliferative capacity, retained adipogenic potential, and upregulated the expression of DNA replication pathway components.
间充质干细胞（MSCs）因其自我更新能力、多向分化潜能和免疫调节作用，是再生治疗的有前景的候选者。MSCs的分子特征受其所处位置的影响。近来，硬膜外脂肪（EF）及其来源的MSCs（EF-MSCs）因其对脊髓微环境的潜在益处以及神经干细胞标志物的高表达而受到关注。然而，由于细胞衰老和EF获取受限，它们的临床应用受到限制。尽管许多研究试图建立永生化、稳定的干细胞系，但永生化EF-MSCs的特征仍有待阐明。
建立并分析稳定的永生化EF-MSCs。
使用光学显微镜分析EF-MSCs的表型。使用慢病毒载体进行细胞永生化。通过免疫印迹、定量PCR和蛋白质组学分析细胞的生物分子特征。
与对照组相比，永生化EF-MSCs的寿命显著延长，成脂潜能和干细胞表面标志物表达得以良好保留。导入人端粒酶逆转录酶基因显著延长了EF-MSCs的寿命。蛋白质组学分析显示，永生化EF-MSCs中DNA复制途径成分的表达大幅增加。
永生化EF-MSCs表现出显著增强的增殖能力，保留了成脂潜能，并上调了DNA复制途径成分的表达。
