V-myc immortalizes human neural stem cells in the absence of pluripotency-associated traits

María José Pino-Barrio; Elisa García-García; Pablo Menéndez; Alberto Martínez-Serrano

doi:10.1371/journal.pone.0118499

V-myc immortalizes human neural stem cells in the absence of pluripotency-associated traits

PLoS One. 2015 Mar 12;10(3):e0118499. doi: 10.1371/journal.pone.0118499. eCollection 2015.

Authors

María José Pino-Barrio¹, Elisa García-García¹, Pablo Menéndez², Alberto Martínez-Serrano¹

Affiliations

¹ Department of Molecular Biology and Center of Molecular Biology "Severo Ochoa" (CBMSO), Universidad Autónoma de Madrid-Consejo Superior de Investigaciones Científicas (UAM-CSIC), Campus UAM Cantoblanco, Madrid, Spain.
² Josep Carreras Leukemia Research Institute and Cell Therapy Program, Facultat de Medicina, University of Barcelona, Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.

Abstract

A better understanding of the molecular mechanisms governing stem cell self-renewal will foster the use of different types of stem cells in disease modeling and cell therapy strategies. Immortalization, understood as the capacity for indefinite expansion, is needed for the generation of any cell line. In the case of v-myc immortalized multipotent human Neural Stem Cells (hNSCs), we hypothesized that v-myc immortalization could induce a more de-differentiated state in v-myc hNSC lines. To test this, we investigated the expression of surface, biochemical and genetic markers of stemness and pluripotency in v-myc immortalized and control hNSCs (primary precursors, that is, neurospheres) and compared these two cell types to human Embryonic Stem Cells (hESCs) and fibroblasts. Using a Hierarchical Clustering method and a Principal Component Analysis (PCA), the v-myc hNSCs associated with their counterparts hNSCs (in the absence of v-myc) and displayed a differential expression pattern when compared to hESCs. Moreover, the expression analysis of pluripotency markers suggested no evidence supporting a reprogramming-like process despite the increment in telomerase expression. In conclusion, v-myc expression in hNSC lines ensures self-renewal through the activation of some genes involved in the maintenance of stem cell properties in multipotent cells but does not alter the expression of key pluripotency-associated genes.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation
Cell Division
Cells, Cultured
Cluster Analysis
Embryonic Stem Cells / cytology
Embryonic Stem Cells / physiology*
Gene Expression Profiling / methods
Gene Expression Regulation*
Genetic Markers / genetics*
Humans
Multipotent Stem Cells / cytology
Multipotent Stem Cells / physiology
Neural Stem Cells / cytology
Neural Stem Cells / physiology*
Oncogene Protein p55(v-myc) / metabolism*
Principal Component Analysis

Substances

Genetic Markers
Oncogene Protein p55(v-myc)

Grants and funding

This work was supported by grants from (to AMS): Spanish Ministry of Economy and Competitiveness (PLE2009–0101, SAF2010–17167), Comunidad Autónoma Madrid (S2011—BMD—2336), Instituto Salud Carlos III (RETICS TerCel, RD12/0019/0013) and European Union (Excell, NMP4—SL—2008–214706); (to PM): Instituto Salud Carlos III (RETICS TerCel, RD12/0019/0006; FIS P110/0449), ERANEt ISCIII—Fondos FEDER (PI12/03112) and the Spanish Association of Cancer Research (CIMEN2011). MJPB was funded by MINECO (PLE2009–0101) and Instituto Salud Carlos III (RETICS TerCel, RD06/0019/0023). This work was also supported by an institutional grant from Fundación Ramón Areces to the Center of Molecular Biology Severo Ochoa. PM lab is supported by: Instituto de Salud Carlos III (ISCIII; E-Rare-2 Call PI12/03112), Ministerio de Economía y Competitividad (MINECO; SAF2013-43065), Generalitat de Catalunya (SGR330) and Obra Social La Caixa-Fundaciò Josep Carreras. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.