肿瘤细胞治疗研究
英文原题：Therapeutic potential of hUC-MSC secretome preconditioned with IFN-γ and/or TNF-α: An in vitro study on Alzheimer's neuronal cell models.
预处理 hUC-MSC 分泌组是阿尔茨海默病一种有前景的治疗策略，因为它能提高神经元细胞活力并促进神经元再生。
阿尔茨海默病是一种进行性神经退行性疾病，特征为毒性淀粉样β蛋白（Aβ）斑块和神经原纤维缠结（NFT）。治疗选择包括人脐带间充质干细胞（hUC-MSC）疗法。其分泌组含有多种有益物质，如可降解Aβ42的脑啡肽酶（CD10）、降低炎症的抗炎细胞因子，以及促进神经元再生的生长因子。本研究旨在使用肿瘤坏死因子α（TNF-α）和/或干扰素γ（IFN-γ）预处理hUC-MSC，以增强这些有益物质的分泌。将hUC-MSC按5×10^3个细胞/cm²接种于T-25培养瓶，在10 mL无异种成分培养基中传代培养。分别仅用TNF-α、仅用IFN-γ及二者联用对hUC-MSC进行预处理。本研究使用10 ng/mL TNF-α和20 ng/mL IFN-γ。预处理48小时后收集分泌组，并通过0.22 μm滤膜过滤。使用神经母细胞瘤SH-SY5Y细胞系进行体外实验，评估分泌组对神经元存活的影响。使用维甲酸（RA）诱导这些细胞分化，再用Aβ42处理以模拟阿尔茨海默病神经元。分泌组使用浓度为5%、10%和20%，以评估神经保护作用。测试了4种分泌组：未经预处理、TNF-α预处理、IFN-γ预处理，以及TNF-α与IFN-γ联合预处理。IFN-γ和TNF-α处理的hUC-MSC中CD10（脑啡肽酶）表达较高，但可溶性脑啡肽酶（sNEP）释放不足。活力结果显示，IFN-γ预处理的分泌组以10%和20%浓度应用，治疗72小时后细胞活力提升最高。TNF-α与IFN-γ联合预处理的分泌组表现出协同效应，尤其在治疗后24和72小时使用5%和10%浓度时。总之，预处理hUC-MSC分泌组有望治疗阿尔茨海默病，可提高神经元细胞活力并促进神经元再生。但仍需进一步研究优化sNEP释放，并在体内模型中提高治疗效力。
Alzheimer's disease is a progressive neurodegenerative disease that is characterized by toxic Amyloid- (A ) plaques and neurofibrillary tangles (NFTs). Treatment options include the use of human umbilical cord mesenchymal stem cell (hUC-MSC)-based therapy. Its secretome contains healing substances such as neprilysin (CD10), which breaks down A 42; anti-inflammatory cytokines, which lower inflammation; and growth factors, which promote neuronal regeneration. The aim of this study was to produce hUC-MSC secretomes preconditioned with tumor necrosis factor-alpha (TNF- ) and/or interferon-gamma (IFN- ) to enhance the secretion of these healing substances. hUC-MSCs were sub-cultured in T-25 flasks at a seeding density of 5 103 cells/cm 2 in 10 mL xeno-free medium. hUC-MSCs were preconditioned with TNF- only, IFN- only, and a combination of TNF- and IFN- . This study used 10 ng/mL TNF- and 20 ng/mL IFN- . The secretome was harvested after 48 hours of preconditioning and then filtered through a 0.22 m filter. In vitro tests were conducted to assess the effects of the secretome on neuronal survival using the neuroblastoma SH-SY5Y cell line. These cells were differentiated with retinoic acid (RA) and then exposed to A 42 to mimic Alzheimer's disease neurons. Secretome therapy was applied at concentrations of 5%, 10%, and 20% to evaluate neuroprotective effects. Four types of secretome were tested: unpreconditioned, TNF- preconditioned, IFN- preconditioned, and a combination of TNF- and IFN- . High levels of CD10 (neprilysin) expression were observed in hUC-MSCs treated with IFN- and TNF- , although they did not release sufficient soluble neprilysin (sNEP). Viability results indicated that secretomes preconditioned with IFN- at 10% and 20% concentrations provided the highest increase in cell viability after 72 hours post-therapy. The combination of TNF- and IFN- preconditioned secretome exhibited synergistic effects, particularly at 5% and 10% doses at 24- and 72-hours post- therapy. In conclusion, preconditioned hUC-MSC secretome represents a promising therapeutic approach for Alzheimer's disease, as it enhances neuronal cell viability and promotes neuronal regeneration. However, further studies are required to optimize sNEP release and maximize therapeutic efficacy in in vivo models.
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