Tondreau Tatiana, Dejeneffe Marielle, Meuleman Nathalie, Stamatopoulos Basile, Delforge Alain, Martiat Philippe, Bron Dominique, Lagneaux Laurence
Institut Jules Bordet, Université Libre de Bruxelles, Laboratory of Experimental Hematology, 121, Bd de Waterloo, 1000 Brussels, Belgium.
BMC Genomics. 2008 Apr 11;9:166. doi: 10.1186/1471-2164-9-166.
Neuronal tissue has limited potential to self-renew or repair after neurological diseases. Cellular therapies using stem cells are promising approaches for the treatment of neurological diseases. However, the clinical use of embryonic stem cells or foetal tissues is limited by ethical considerations and other scientific problems. Thus, bone marrow mesenchymal stomal cells (BM-MSC) could represent an alternative source of stem cells for cell replacement therapies. Indeed, many studies have demonstrated that MSC can give rise to neuronal cells as well as many tissue-specific cell phenotypes.
BM-MSC were differentiated in neuron-like cells under specific induction (NPBM + cAMP + IBMX + NGF + Insulin). By day ten, differentiated cells presented an expression profile of real neurons. Functionality of these differentiated cells was evaluated by calcium influx through glutamate receptor AMPA3.
Using microarray analysis, we compared gene expression profile of these different samples, before and after neurogenic differentiation. Among the 1943 genes differentially expressed, genes down-regulated are involved in osteogenesis, chondrogenesis, adipogenesis, myogenesis and extracellular matrix component (tuftelin, AGC1, FADS3, tropomyosin, fibronectin, ECM2, HAPLN1, vimentin). Interestingly, genes implicated in neurogenesis are increased. Most of them are involved in the synaptic transmission and long term potentialisation as cortactin, CASK, SYNCRIP, SYNTL4 and STX1. Other genes are involved in neurite outgrowth, early neuronal cell development, neuropeptide signaling/synthesis and neuronal receptor (FK506, ARHGAP6, CDKRAP2, PMCH, GFPT2, GRIA3, MCT6, BDNF, PENK, amphiregulin, neurofilament 3, Epha4, synaptotagmin). Using real time RT-PCR, we confirmed the expression of selected neuronal genes: NEGR1, GRIA3 (AMPA3), NEF3, PENK and Epha4. Functionality of these neuron-like cells was demonstrated by Ca2+ influx through glutamate receptor channel (AMPA3) in the presence of two agonist glutamate, AMPA or CNQX antagonist.
Our results demonstrate that BM-MSC have the potential to differentiate in neuronal cells with specific gene expression and functional properties. BM-MSC are thus promising candidates for cell-based therapy of neurodegenerative diseases.
神经组织在神经疾病后自我更新或修复的潜力有限。使用干细胞的细胞疗法是治疗神经疾病的有前景的方法。然而，胚胎干细胞或胎儿组织的临床应用受到伦理考量和其他科学问题的限制。因此，骨髓间充质基质细胞（BM-MSC）可能是细胞替代疗法中干细胞的另一种来源。事实上，许多研究已证明间充质干细胞可分化为神经元细胞以及许多组织特异性细胞表型。
在特定诱导条件下（NPBM + cAMP + IBMX + NGF + 胰岛素）将BM-MSC分化为类神经元细胞。到第10天时，分化细胞呈现出真正神经元的表达谱。通过谷氨酸受体AMPA3介导的钙内流来评估这些分化细胞的功能。
使用微阵列分析，我们比较了神经源性分化前后这些不同样本的基因表达谱。在1943个差异表达基因中，下调的基因参与成骨、软骨形成、脂肪形成、肌生成和细胞外基质成分（牙本质磷蛋白、AGC1、FADS3、原肌球蛋白、纤连蛋白、ECM2、HAPLN1、波形蛋白）。有趣的是，与神经发生相关的基因增加。其中大多数参与突触传递和长期增强作用，如皮层肌动蛋白、CASK、SYNCRIP、SYNTL4和STX1。其他基因参与神经突生长、早期神经元细胞发育、神经肽信号传导/合成和神经元受体（FK506、ARHGAP6、CDKRAP2、PMCH、GFPT2、GRIA3、MCT6、BDNF、PENK、双调蛋白、神经丝3、Epha4、突触结合蛋白）。使用实时RT-PCR，我们证实了所选神经元基因的表达：NEGR1、GRIA3（AMPA3）、NEF3、PENK和Epha4。在存在两种激动剂谷氨酸、AMPA或CNQX拮抗剂的情况下，通过谷氨酸受体通道（AMPA3）介导的Ca2+内流证明了这些类神经元细胞的功能。
我们的结果表明BM-MSC有潜力分化为具有特定基因表达和功能特性的神经元细胞。因此，BM-MSC是神经退行性疾病细胞治疗的有前景的候选者。
