Agner Henrique Dorigo Hochuli, Ana Helena Selenko, Mateus de Oliveira Lisboa, Alexandra Cristina Senegaglia, Maria Luiza de Castro, Joselito Getz, Letícia Fracaro, Paulo Roberto Slud Brofman
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引用次数: 0
Abstract
Introduction: Mesenchymal stromal cells (MSCs) play a crucial role in tissue repair and exhibit anti-inflammatory properties, making them promising for regenerative medicine. Dental tissue-derived MSCs, such as stromal cells from human exfoliated deciduous teeth (SHEDs) and dental pulp stromal cells (DPSCs), express neural markers and hold the potential for treating neurodegenerative diseases. Nonetheless, their relative ability to differentiate into neurons is still unclear. Given their shared embryonic origin, we hypothesised that SHEDs and DPSCs possess similar potential for neuronal differentiation along with intrinsic expression of neuronal markers. The objective of this study was to compare their differentiation abilities under standardised conditions, evaluate neuronal markers pro- and post-neuronal induction, and compare the neuronal differentiation potential of SHEDs and DPSCs.
Methods: SHEDs (n = 3) and DPSCs (n = 3) were collected with ethical approval, cultured, and characterised according to established MSC criteria. Clonogenicity, proliferation, senescence, and trilineage differentiation were assessed. Neuronal differentiation was induced for 21 days and evaluated using flow cytometry (SRY-Box Transcription Factor 1 [SOX1], SRY-Box Transcription Factor 2 [SOX2], glial fibrillary acidic protein [GFAP], doublecortin, nestin, CD56, CD146), immunofluorescence for βIII-tubulin, reverse transcription polymerase chain reaction for tubulin 3 (TUB3), and microtubule-associated protein 2 (MAP2).
Results: Both SHEDs and DPSCs exhibited MSC characteristics. SHEDs showed higher clonogenicity. Early neuronal markers (e.g., SOX1, nestin, GFAP, βIII-tubulin) were detected pre- and post-induction in both cell types without significant intergroup differences. No significant expression of TUB3 and MAP2 was observed.
Conclusion: SHEDs and DPSCs show comparable neuronal marker expression profiles, suggesting similar early neuronal differentiation potential. These findings support using undifferentiated SHEDs and DPSCs in neuroregenerative strategies, offering cost-effective and safer alternatives to pre-differentiated cells.
间充质基质细胞(MSCs)在组织修复中起着至关重要的作用,并具有抗炎特性,使其在再生医学中具有广阔的应用前景。牙组织来源的间质干细胞,如来自人脱落乳牙(SHEDs)和牙髓间质细胞(DPSCs)的间质细胞,表达神经标记物,具有治疗神经退行性疾病的潜力。尽管如此,它们分化为神经元的相对能力仍不清楚。鉴于它们共同的胚胎起源,我们假设shed和DPSCs具有相似的神经元分化潜力以及神经元标记物的内在表达。本研究的目的是比较它们在标准化条件下的分化能力,并评估神经元诱导前后的神经元标志物。目的:比较shed和DPSCs的神经元分化潜能。方法:收集经伦理批准的牛舍(n=3)和DPSCs (n=3),根据既定的MSC标准进行培养和表征。克隆原性、增殖、衰老和三龄分化进行了评估。诱导神经元分化21天,采用流式细胞术(SRY-Box转录因子1 (SOX1)、SRY-Box转录因子2 (SOX2)、胶质纤维酸性蛋白(GFAP)、双皮质素、巢蛋白、CD56、CD146)、免疫荧光法检测β iii -微管蛋白,反转录聚合酶链反应(RT-PCR)检测微管蛋白3 (TUB3)和微管相关蛋白2 (MAP2)。结果:shed和DPSCs均表现出MSC特征。棚舍表现出较高的克隆原性。在两种细胞类型诱导前和诱导后均检测到早期神经元标志物(如SOX1、Nestin、GFAP、β iii -微管蛋白),组间差异不显著。TUB3和MAP2未见明显表达。结论:shed和DPSCs具有相似的神经元标志物表达谱,提示相似的早期神经元分化潜力。这些发现支持在神经再生策略中使用未分化的shed和DPSCs,为预分化细胞提供了成本效益和更安全的替代品。
期刊介绍:
''Cells Tissues Organs'' aims at bridging the gap between cell biology and developmental biology and the emerging fields of regenerative medicine (stem cell biology, tissue engineering, artificial organs, in vitro systems and transplantation biology). CTO offers a rapid and fair peer-review and exquisite reproduction quality. Special topic issues, entire issues of the journal devoted to a single research topic within the range of interests of the journal, are published at irregular intervals.