BACKGROUND Spinal cord injury (SCI) is a complex and severe neurological condition. Mesenchymal stem cells (MSCs) and their secreted factors show promising potential for regenerative medicine. Many studies have investigated MSC expansion efficacy of all kinds of culture medium formulations, such as growth factor-supplemented or xeno-free medium. However, very few studies have focused on the potential of human MSC (hMSC) culture medium formulations for injured spinal cord repair. In this study, we investigated the effect of hMSC-conditioned medium supplemented with bFGF, EGF, and patient plasma, namely, neural regeneration laboratory medium (NRLM), on SCI in vitro and in vivo. METHODS Commercial and patient bone marrow hMSCs were obtained for cultivation in standard medium and NRLM separately. Several characteristics, including CD marker expression, differentiation, and growth curves, were compared between MSCs cultured in standard medium and NRLM. Additionally, we investigated the effect of the conditioned medium (referred to as NRLM-CM) on neural repair, including inflammation inhibition, neurite regeneration, and spinal cord injury (SCI), and used a coculture system to detect the neural repair function of NRLM-MSCs. RESULTS Compared to standard culture medium, NRLM-CM had superior in inflammation reduction and neurite regeneration effects in vitro and improved functional restoration in SCI rats in vivo. In comparison with standard culture medium MSCs, NRLM-MSCs proliferated faster regardless of the age of the donor. NRLM-MSCs also showed increased adipose differentiative potential and reduced CD90 expression. Both types of hMSC CM effectively enhanced injured neurite outgrowth and protected against H2O2 toxicity in spinal cord neuron cultures. Cytokine arrays performed in hMSC-CM further revealed the presence of at least 120 proteins. Among these proteins, 6 demonstrated significantly increased expression in NRLM-CM: adiponectin (Acrp30), angiogenin (ANG), HGF, NAP-2, uPAR, and IGFBP2. CONCLUSIONS The NRLM culture system provides rapid expansion effects and functional hMSCs. The superiority of the derived conditioned medium on neural repair shows potential for future clinical applications.

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来源于快速扩增的间充质干细胞的条件培养基对神经修复的优越性。

背景技术脊髓损伤（SCI）是一种复杂而严重的神经系统疾病。间充质干细胞（MSCs）及其分泌的因子显示出有希望的再生医学潜力。许多研究已经研究了各种培养基制剂（例如补充生长因子或无异种培养基）的MSC扩增功效。但是，很少有研究集中在人类MSC（hMSC）培养基制剂对受损脊髓修复的潜力上。在这项研究中，我们调查了补充bFGF，EGF和患者血浆的hMSC条件培养基（即神经再生实验室培养基（NRLM））在体外和体内对SCI的影响。方法获得商业和患者骨髓hMSCs，分别在标准培养基和NRLM中培养。几个特点 比较了在标准培养基和NRLM中培养的MSC之间的CD标记表达，分化和生长曲线。此外，我们研究了条件培养基（称为NRLM-CM）对神经修复的影响，包括炎症抑制，神经突再生和脊髓损伤（SCI），并使用共培养系统检测NRLM的神经修复功能-MSC。结果与标准培养基相比，NRLM-CM具有更好的体外消炎和神经突再生作用，并能改善SCI大鼠体内的功能恢复。与标准培养基MSC相比，NRLM-MSC的增殖速度更快，而与供体的年龄无关。NRLM-MSCs还显示出增加的脂肪分化潜能和减少的CD90表达。两种类型的hMSC CM在脊髓神经元培养物中均能有效增强受伤的神经突向外生长并防止H2O2毒性。在hMSC-CM中进行的细胞因子阵列进一步揭示了至少120种蛋白质的存在。在这些蛋白质中，有6种在NRLM-CM中表现出明显增加的表达：脂联素（Acrp30），血管生成素（ANG），HGF，NAP-2，uPAR和IGFBP2。结论NRLM培养系统具有快速扩增作用和功能性hMSC。衍生条件培养基对神经修复的优越性显示了其在未来临床应用中的潜力。脂联素（Acrp30），血管生成素（ANG），HGF，NAP-2，uPAR和IGFBP2。结论NRLM培养系统具有快速扩增作用和功能性hMSC。衍生条件培养基对神经修复的优越性显示了其在未来临床应用中的潜力。脂联素（Acrp30），血管生成素（ANG），HGF，NAP-2，uPAR和IGFBP2。结论NRLM培养系统具有快速扩增作用和功能性hMSC。衍生条件培养基对神经修复的优越性显示了其在未来临床应用中的潜力。