Department of Histology and Embryology, İstanbul University-Cerrahpaşa, Cerrahpaşa Faculty of Medicine, İstanbul, Türkiye.
Department of Histology and Embryology, Balıkesir University Faculty of Medicine, Balıkesir, Türkiye.
Balkan Med J. 2024 Sep 6;41(5):369-376. doi: 10.4274/balkanmedj.galenos.2024.2024-6-25.
Mesenchymal stem cells (MSCs) play a key role in regenerative medicine due to their capacity to differentiate into multiple cell lines, regulate the immune system, and exert paracrine effects. The therapeutic impact of MSCs is primarily mediated through their secretome. The secretory and therapeutic potential of MSCs can be improved through preconditioning, which entails the application of hypoxic environments, 3-dimensional cell cultures, and pharmacological agents. Valproic acid (VPA) is a histone deacetylase inhibitor that is employed in medical practice for treating epilepsy and bipolar disorder. Hence, preconditioning MSCs with VPA is expected to induce histone acetylation, enhance gene expression, and beneficially modify the cells' secretomes.
To assess the effectiveness of VPA in enhancing and regulating the therapeutic potential of cells as well as its impact on MSC secretome profiles and ultrastructural morphologies.
Expiremental study.
Human umbilical cord MSCs were preconditioned with 2 mM VPA for 24 and 48 hours; untreated MSCs served as controls. The secretome secreted by the cells was assessed for its total protein content. Subsequently, interferon-gamma (IFN-γ), interleukin-17 (IL-17), IL-10, vascular endothelial growth factor, nerve growth factor (NGF), glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor (BDNF) levels in the secretome were analyzed using the ELISA method. The ultrastructural properties of the cells were studied under transmission electron microscopy.
Ultrastructural examinations revealed that the chromatin content of VPA-treated cells was reduced. VPA-preconditioned cells exhibited a higher density of rough endoplasmic reticulum, autophagic vesicles, and myelin figures on cytoplasmic structure analysis, which was indicative of increased secretion. Protein secretion was elevated in those cells, with notable increases in NGF and BDNF levels. Furthermore, the cytoskeletal rearrangement and elevated autophagic activity observed in the 48-hour preconditioned cells could indicate the initiation of neuronal differentiation. IL-10, IL-17, and IFN-γ were not detected in the secretome.
This study indicate that preconditioning with VPA enhances MSC activity and subsequently modifies the secretome content.
间充质干细胞（MSCs）因其能够分化为多种细胞系、调节免疫系统和发挥旁分泌作用而在再生医学中发挥关键作用。MSCs 的治疗作用主要通过其分泌组来介导。通过预处理可以改善 MSCs 的分泌和治疗潜力，预处理包括应用低氧环境、三维细胞培养和药物制剂。丙戊酸（VPA）是一种组蛋白去乙酰化酶抑制剂，在医学实践中用于治疗癫痫和双相情感障碍。因此，用 VPA 预处理 MSCs 有望诱导组蛋白乙酰化、增强基因表达，并有益地修饰细胞的分泌组。
评估 VPA 增强和调节细胞治疗潜力的效果及其对 MSC 分泌组谱和超微结构形态的影响。
实验研究。
用 2mM VPA 预处理人脐带 MSC 24 小时和 48 小时；未处理的 MSC 作为对照。评估细胞分泌的分泌组中的总蛋白含量。随后，使用 ELISA 法分析分泌组中干扰素-γ（IFN-γ）、白细胞介素-17（IL-17）、IL-10、血管内皮生长因子、神经生长因子（NGF）、胶质细胞源性神经营养因子和脑源性神经营养因子（BDNF）的水平。通过透射电子显微镜研究细胞的超微结构特性。
超微结构检查显示，VPA 处理细胞的染色质含量减少。VPA 预处理细胞的细胞质结构分析显示，粗面内质网、自噬小体和髓鞘结构的密度增加，表明分泌增加。这些细胞的蛋白质分泌增加，NGF 和 BDNF 水平显著升高。此外，在 48 小时预处理的细胞中观察到细胞骨架重排和自噬活性增加，这可能表明开始向神经元分化。在分泌组中未检测到 IL-10、IL-17 和 IFN-γ。
本研究表明，VPA 预处理可增强 MSC 活性，随后改变分泌组内容。
