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Differential pathotropism of non-immortalized and immortalized human neural stem cell lines in a focal demyelination model

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Abstract

Cell therapy is reaching the stage of phase I clinical trials for post-traumatic, post-ischemic, or neurodegenerative disorders, and the selection of the appropriate cell source is essential. In order to assess the capacity of different human neural stem cell lines (hNSC) to contribute to neural tissue regeneration and to reduce the local inflammation after an acute injury, we transplanted GMP-grade non-immortalized hNSCs and v-myc (v-IhNSC), c-myc T58A (T-IhNSC) immortalized cells into the corpus callosum of adult rats after 5 days from focal demyelination induced by lysophosphatidylcholine. At 15 days from transplantation, hNSC and T-IhNSC migrated to the lesioned area where they promoted endogenous remyelination and differentiated into mature oligodendrocytes, while the all three cell lines were able to integrate in the SVZ. Moreover, where demyelination was accompanied by an inflammatory reaction, a significant reduction of microglial cells’ activation was observed. This effect correlated with a differential migratory pattern of transplanted hNSC and IhNSC, significantly enhanced in the former, thus suggesting a specific NSC-mediated immunomodulatory effect on the local inflammation. We provide evidence that, in the subacute phase of a demyelination injury, different human immortalized and non-immortalized NSC lines, all sharing homing to the stem niche, display a differential pathotropism, both through cell-autonomous and non-cell autonomous effects. Overall, these findings promote IhNSC as an inexhaustible cell source for large-scale preclinical studies and non-immortalized GMP grade hNSC lines as an efficacious, safe, and reliable therapeutic tool for future clinical applications.

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Acknowledgments

We thank Pietro De Filippis, Patrizia Karoschitz, Cesare Rota Nodari, Loredana Turani, Antonio Tomaino, for precious suggestions. This work was supported by the CARIPLO Foundation; Neurothon ONLUS Foundation; Italian Association for Cancer Research (AIRC); Italian Ministry of Health (Ricerca Finalizzata). This work was financially supported by grants from the CARIPLO Foundation, the Neurothon ONLUS Foundation, Fondazione Borgonovo ONLUS, and by the Italian Association for Cancer Research (AIRC), the Italian Ministry of Health (Ricerca Finalizzata), and Stemgen S.p.a.

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Authors and Affiliations

  1. Department of Biotechnology and Biosciences, Università Milano Bicocca, Piazza della Scienza 2, 20126, Milan, Italy

    Daniela Ferrari, Cristina Zalfa, Laura Rota Nodari, Angelo Luigi Vescovi & Lidia De Filippis

  2. Casa Sollievo Della Sofferenza, S. Giovanni Rotondo (FG), Italy

    Maurizio Gelati & Angelo Luigi Vescovi

  3. Laboratorio Cellule Staminali, Cell Factory e Biobanca, Azienda ospedaliera “Santa Maria”, Terni, Italy

    Maurizio Gelati & Angelo Luigi Vescovi

  4. Department of Experimental Oncology, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy

    Luigi Carlessi & Domenico Delia

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  1. Daniela Ferrari
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  2. Cristina Zalfa
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  7. Angelo Luigi Vescovi
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Correspondence to Daniela Ferrari or Lidia De Filippis.

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D. Ferrari and C. Zalfa contributed equally to this work.

The present work was developed at the University of Milano-Bicocca, Department of Biotechnologies and Biosciences, Milan, Italy.

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18_2011_873_MOESM1_ESM.tif

Supplementary Fig. 1 Differential expression of CXCL12 and EGF in the lesioned versus contralateral hemisphere. Confocal images showing the enhanced expression of CXCL12 (A–B) and EGF (C–D), respectively, in the lesioned (A and C) versus the contralateral (B and D) hemisphere. Total nuclei are shown by DAPI staining (blue). Scale bars = 75 μm (TIFF 30871 kb)

18_2011_873_MOESM2_ESM.tif

Supplementary Fig. 2 Coimmunolabeled huNestin +/GFAP + cells in transplanted animals. Confocal analysis of huNestin + (green)/GFAP + (red) cells along the SVZ wall deriving from hNSC. Total nuclei are shown by DAPI staining (blue). Scale bars = 75 μm (inset = 14 μm). lv lateral ventricle, cc corpus callosum (TIFF 3368 kb)

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Ferrari, D., Zalfa, C., Rota Nodari, L. et al. Differential pathotropism of non-immortalized and immortalized human neural stem cell lines in a focal demyelination model. Cell. Mol. Life Sci. 69, 1193–1210 (2012). https://doi.org/10.1007/s00018-011-0873-5

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