Department of Neurology and Laboratory of Neuroscience, IRCCS Istituto Auxologico Italiano, Cusano Milanino 20095, Italy.
Neuroscience. 2013 Mar 27;234:158-72. doi: 10.1016/j.neuroscience.2012.12.038. Epub 2013 Jan 3.
The capability to integrate into degenerative environment, release neurotrophic cytokines, contrast oxidative stress and an inherent differentiation potential towards siteappropriate phenotypes are considered crucial for the use of stem cells in tissue repair and regeneration. Naïve human chorial villi- (hCVCs) and amniotic fluid- (hAFCs) derived cells, whose properties and potentiality have not been extensively investigated, may represent two novel foetal cell sources for stem cell therapy. We previously described that long-term transplantation of hAFCs in the lateral ventricles of wobbler and healthy mice was feasible and safe. In the present study we examine the in vitro intrinsic stem potential of hCVCs and hAFCs for future therapeutic applications in neurodegenerative disorders. Presence of stem lineages was evaluated assessing the expression pattern of relevant candidate markers by flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Release of cytokines that may potentialy sustain endogenous neurogenesis and/or activate neuroprotective pathways was quantified by enzyme-linked immunosorbent assays (ELISAs). We also performed an in vitro neurorescue assay, wherein a neuroblastoma cell line damaged by 6-hydroxydopamine (6-OHDA) was treated with hCVC/hAFC-derived conditioned medium (CM). Naïve hCVCs/hAFCs show a neurogenic/angiogenic predisposition. Both cell types express several specific neural stem/progenitor markers, such as nestin and connexin 43, and release significant amounts of brain-derived neurotrophic factor, as well as vascular endothelial growth factor. hCVC and hAFC populations comprise several interesting cell lineages, including mesenchymal stem cells (MSCs) and cells with neural-like phenotypes. Moreover, although CMs obtained from both cell cultures actively sustained metabolic activity in a 6-OHDA-induced Parkinson's disease (PD) cell model, only hCVC-derived CMs significantly reduced neurotoxin-induced apoptosis. In conclusion, this study demonstrates that naïve hAFCs and hCVCs may enhance cell-recovery following neuronal damage through multiple rescue mechanisms, and may provide a suitable means of stem cell therapy for neurodegenerative disorders including PD.
能够适应退化环境、释放神经营养因子、对抗氧化应激以及向特定部位分化的固有潜能，这些被认为是将干细胞应用于组织修复和再生的关键。尚未广泛研究其特性和潜能的原始人绒毛膜绒毛（hCVCs）和羊水（hAFCs）衍生细胞，可能代表了干细胞治疗的两种新型胎儿细胞来源。我们之前曾描述过，将 hAFC 长期移植到 wobbler 和健康小鼠的侧脑室是可行且安全的。在本研究中，我们检查了 hCVC 和 hAFC 的内在干细胞潜能，以用于神经退行性疾病的未来治疗应用。通过流式细胞术、逆转录-聚合酶链反应（RT-PCR）和免疫细胞化学评估相关候选标志物的表达模式，来评估干细胞谱系的存在。通过酶联免疫吸附试验（ELISA）定量测定可能维持内源性神经发生和/或激活神经保护途径的细胞因子释放。我们还进行了体外神经挽救测定，其中用 6-羟多巴胺（6-OHDA）损伤的神经母细胞瘤细胞系用 hCVC/hAFC 衍生的条件培养基（CM）处理。原始 hCVC/hAFC 具有神经发生/血管生成倾向。这两种细胞类型都表达几种特定的神经干/祖细胞标志物，如巢蛋白和连接蛋白 43，并释放大量脑源性神经营养因子和血管内皮生长因子。hCVC 和 hAFC 群体包含多种有前途的细胞谱系，包括间充质干细胞（MSCs）和具有神经样表型的细胞。此外，尽管两种细胞培养物的 CM 均积极维持 6-OHDA 诱导的帕金森病（PD）细胞模型中的代谢活性，但只有 hCVC 衍生的 CM 可显著减少神经毒素诱导的细胞凋亡。总之，这项研究表明，原始 hAFCs 和 hCVCs 可以通过多种挽救机制增强神经元损伤后的细胞恢复，并可能为包括 PD 在内的神经退行性疾病提供合适的干细胞治疗手段。
