Neurogenic potential of human mesenchymal stem cells revisited: analysis by immunostaining, time-lapse video and microarray
Author:
Bertani Nicoletta1, Malatesta Paolo1, Volpi Giorgia1, Sonego Paolo2, Perris Roberto12
Affiliation:
1. Department of Evolutionary and Functional Biology, University of Parma, Viale delle Scienze 11/a, 43100 Parma, Italy 2. Division for Experimental Oncology 2, The National Cancer Institute, CRO-IRCCS, Via Pedemontana Occidentale 1, Aviano 33081, Italy
Abstract
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.
Publisher
The Company of Biologists
Reference62 articles.
1. Alison, M. R., Poulsom, R., Jeffery, R., Dhillon, A. P., Quaglia, A., Jacob, J., Novelli, M., Prentice, G., Williamson, J. and Wright, N. A. (2000). Hepatocytes from non-hepatic adult stem cells. Nature406, 257. 2. Alvarez-Dolado, M., Pardal, R., Garcia-Verdugo, J. M., Fike, J. R., Lee, H. O., Pfeffer, K., Lois, C., Morrison, S. J. and Alvarez-Buylla, A. (2003). Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature425, 968-973. 3. Blau, H. M., Brazelton, T. R. and Weimann, J. M. (2001). The evolving concept of a stem cell: entity or function? Cell105, 829-841. 4. Colter, D. C., Sekiya, I. and Prockop, D. J. (2001). Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proc. Natl. Acad. Sci. USA98, 7841-7845. 5. Conget, P. A. and Minguell, J. J. (1999). Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J. Cell Physiol.181, 67-73.
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