Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, 11221, Taiwan.
Neural Regeneration Laboratory, Department of Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, 11217, Taiwan.
Stem Cell Res Ther. 2019 Dec 16;10(1):390. doi: 10.1186/s13287-019-1491-7.
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.
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.
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 HO 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.
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.
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) 及其分泌因子在再生医学中显示出有希望的潜力。许多研究已经调查了各种培养基配方的 MSC 扩增效果，例如生长因子补充或无动物源培养基。然而，很少有研究关注人 MSC (hMSC) 培养基配方对损伤脊髓修复的潜力。在这项研究中，我们研究了补充 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)，对 SCI 的体外和体内作用。
分别从商业来源和患者骨髓中获得 hMSC 进行培养，分别在标准培养基和 NRLM 中培养。比较了在标准培养基和 NRLM 中培养的 MSC 的几个特征，包括 CD 标志物表达、分化和生长曲线。此外，我们研究了条件培养基 (称为 NRLM-CM) 对神经修复的作用，包括抑制炎症、促进神经突再生和脊髓损伤 (SCI)，并使用共培养系统检测 NRLM-MSCs 的神经修复功能。
与标准培养相比，NRLM-CM 在体外具有更好的减少炎症和促进神经突再生的作用，并在体内改善 SCI 大鼠的功能恢复。与标准培养的 MSC 相比，无论供体年龄如何，NRLM-MSC 增殖更快。NRLM-MSC 还显示出增强的脂肪分化潜能和降低的 CD90 表达。两种类型的 hMSC CM 均能有效增强受损神经突的生长，并防止脊髓神经元培养物中 HO 毒性。在 hMSC-CM 中进行的细胞因子阵列进一步显示存在至少 120 种蛋白质。在这些蛋白质中，有 6 种在 NRLM-CM 中表达显著增加：脂联素 (Acrp30)、血管生成素 (ANG)、HGF、NAP-2、uPAR 和 IGFBP2。
NRLM 培养系统提供快速扩增效果和功能 hMSC。衍生的条件培养基在神经修复方面的优越性显示出未来临床应用的潜力。
