Department of Neuroscience, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, v.v.i., Videnska 1083, 142 20 Prague 4, Czech Republic.
Cell Mol Neurobiol. 2011 Oct;31(7):1113-22. doi: 10.1007/s10571-011-9712-3. Epub 2011 Jun 1.
Adipose-derived stromal cells (ASCs) are an alternative source of stem cells for cell-based therapies of neurological disorders such as spinal cord injury (SCI). In the present study, we predifferentiated ASCs (pASCs) and compared their behavior with naïve ASCs in vitro and after transplantation into rats with a balloon-induced compression lesion. ASCs were predifferentiated into spheres before transplantation, then pASCs or ASCs were injected intraspinally 1 week after SCI. The cells' fate and the rats' functional outcome were assessed using behavioral, histological, and electrophysiological methods. Immunohistological analysis of pASCs in vitro revealed the expression of NCAM, NG2, S100, and p75. Quantitative RT-PCR at different intervals after neural induction showed the up-regulated expression of the glial markers NG2 and p75 and the neural precursor markers NCAM and Nestin. Patch clamp analysis of pASCs revealed three different types of membrane currents; however, none were fast activating Na(+) currents indicating a mature neuronal phenotype. Significant improvement in both the pASC and ASC transplanted groups was observed in the BBB motor test. In vivo, pASCs survived better than ASCs did and interacted closely with the host tissue, wrapping host axons and oligodendrocytes. Some transplanted cells were NG2- or CD31-positive, but no neuronal markers were detected. The predifferentiation of ASCs plays a beneficial role in SCI repair by promoting the protection of denuded axons; however, functional improvements were comparable in both the groups, indicating that repair was induced mainly through paracrine mechanisms.
脂肪来源的基质细胞（ASCs）是一种替代的干细胞来源，可用于治疗神经退行性疾病，如脊髓损伤（SCI）的细胞治疗。在本研究中，我们对 ASC 进行预分化（pASC），并在体外和移植到气囊诱导压迫损伤大鼠后，将其与未分化的 ASC 进行了比较。在移植前，将 ASC 预分化为球体，然后在 SCI 后 1 周将 pASC 或 ASC 注射到脊髓内。使用行为学、组织学和电生理学方法评估细胞的命运和大鼠的功能结果。体外对 pASC 的免疫组织化学分析显示 NCAM、NG2、S100 和 p75 的表达。神经诱导后不同时间点的定量 RT-PCR 显示神经胶质标志物 NG2 和 p75 以及神经前体细胞标志物 NCAM 和 Nestin 的表达上调。pASC 的膜片钳分析显示三种不同类型的膜电流；然而，没有快速激活的 Na(+)电流，表明其具有成熟神经元表型。在 BBB 运动测试中，pASC 和 ASC 移植组均观察到明显的改善。在体内，pASC 的存活率高于 ASC，与宿主组织密切相互作用，包裹宿主轴突和少突胶质细胞。一些移植细胞为 NG2 或 CD31 阳性，但未检测到神经元标志物。ASC 的预分化在 SCI 修复中发挥有益作用，通过促进裸露轴突的保护来促进修复；然而，两组的功能改善相当，表明修复主要是通过旁分泌机制诱导的。
