Neurogenic potential of human mesenchymal stem cells revisited: analysis by immunostaining, time-lapse video and microarray
Neurogenic potential of human mesenchymal stem cells revisited: analysis by immunostaining, time-lapse video and microarray
复制标题
DOI：
10.1242/jcs.02511
复制
发表时间：
2005-09-01
影响因子：
4
通讯作者：
Perris, R
中科院分区：
文献类型：
--
作者：
Bertani, N;Malatesta, P;Perris, R
通过简单的体外处理从人骨髓间充质干细胞（hMSCs）产生神经细胞的可能性在概念上和实践上都很有吸引力。然而，这种干细胞的化学操作后观察到的表型调制是否真正代表了真正的跨谱系分化仍有待建立。我们已经重新评估了一个经常报道的生化方法的影响，基于治疗与丁基羟基茴香醚和二甲亚砜，带来这样的表型转换，通过监测的形态变化引起的治疗在真实的时间，通过分析表型特异性蛋白质标记物的表达和评估的调制转录。视频延时显微镜显示，间充质干细胞向神经元样形态的转化可以再现。在正常的原代成纤维细胞中，以及通过添加引起细胞骨架塌陷和局部粘附接触破坏的药物来模拟。标记物分析显示，间充质干细胞组成性表达多谱系特征，包括几个有关的神经之一。然而，所应用的“神经诱导”方案既不显著调节这些标记物的表达，也不诱导其他神经特异性蛋白质的从头翻译。同样，超过21，000个基因的全球表达谱表明基因转录受到的影响很小。最引人注目的是，我们发现，诱导治疗改变表达的基因组与比较未处理的间充质干细胞和未成熟神经组织时差异表达的基因组不匹配。相反，通过将这些基因表达谱与从相同细胞和不相关的非神经器官（如肝脏）之间的比较中获得的基因表达谱进行比较，我们发现所采用的神经诱导方案在将人类间充质干细胞重定向到神经表型方面并不比重定向到内胚层肝脏途径更有效。
The possibility of generating neural cells from human bone-marrow-derived mesenchymal stem cells (hMSCs) by simple in vitro treatments is appealing both conceptually and practically. However, whether phenotypic modulations observed after chemical manipulation of such stem cells truly represent a genuine trans-lineage differentiation remains to be established. We have re-evaluated the effects of a frequently reported biochemical approach, based on treatment with butylated hydroxyanisole and dimethylsulphoxide, to bring about such phenotypic conversion by monitoring the morphological changes induced by the treatment in real time, by analysing the expression of phenotype-specific protein markers and by assessing the modulation of transcriptome. Video time-lapse microscopy showed that conversion of mesenchymal stem cells to a neuron-like morphology could be reproduced. in normal primary fibroblasts as well as mimicked by addition of drugs eliciting cytoskeletal collapse and disruption of focal adhesion contacts. Analysis of markers revealed that mesenchymal stem cells constitutively expressed multi-lineage traits, including several pertaining to the neural one. However, the applied 'neural induction' protocol neither significantly modulated the expression of such markers, nor induced de novo translation of other neural-specific proteins. Similarly, global expression profiling of over 21,000 genes demonstrated that gene transcription was poorly affected. Most strikingly, we found that the set of genes whose expression was altered by the inductive treatment did not match those sets of genes differentially expressed when comparing untreated mesenchymal stem cells and immature neural tissues. Conversely, by comparing these gene expression profiles with that obtained from comparisons between the same cells and an unrelated non-neural organ, such as liver, we found that the adopted neural induction protocol was no more effective in redirecting human mesenchymal stem cells toward a neural phenotype than toward an endodermal hepatic pathway.