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In vitro characterization of subventricular zone isolated neural stem cells, from adult monkey and rat brain

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Abstract

Neural stem cells (NSCs) are multipotent, self-renewable cells who are capable of differentiating into neurons, astrocytes, and oligodendrocytes. NSCs reside at the subventricular zone (SVZ) of the adult brain permanently to guarantee a lifelong neurogenesis during neural network plasticity or undesirable injuries. Although the specious inaccessibility of adult NSCs niche hampers their in vivo identification, researchers have been seeking ways to optimize adult NSCs isolation, expansion, and differentiation, in vitro. NSCs were isolated from rhesus monkey SVZ, expanded in vitro and then characterized for NSCs-specific markers expression by immunostaining, real-time PCR, flow cytometry, and cell differentiation assessments. Moreover, cell survival as well as self-renewal capacity were evaluated by TUNEL, Live/Dead and colony assays, respectively. In the next step, to validate SVZ-NSCs identity in other species, a similar protocol was applied to isolate NSCs from adult rat’s SVZ as well. Our findings revealed that isolated SVZ-NSCs from both monkey and rat preserve proliferation capacity in at least nine passages as confirmed by Ki67 expression. Additionally, both SVZ-NSCs sources are capable of self-renewal in addition to NESTIN, SOX2, and GFAP expression. The mortality was measured meager with over 95% viability according to TUNEL and Live/Dead assay results. Eventually, the multipotency of SVZ-NSCs appraised authentic after their differentiation into neurons, astrocytes, and oligodendrocytes. In this study, we proposed a reliable method for SVZ-NSCs in vitro maintenance and identification, which, we believe is a promising cell source for therapeutic approach to recover neurological disorders and injuries condition.

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Funding

The present research was completely funded by the Iranian Council of Lotus. This research involved the three Rhesus monkeys and three Wistar rats that were cared, fed and behaved under ethical standards.

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Authors and Affiliations

  1. Department of Stem Cell and Developmental Biology, Cell Science Research Center, ROYAN Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Razieh Jaberi, Sara Mirsadeghi & Sahar Kiani

  2. Department of Brain and Cognitive Sciences, Cell Science Research Center, ROYAN Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Razieh Jaberi, Sara Mirsadeghi & Sahar Kiani

  3. Department of Developmental Biology, University of Science and Culture, Tehran, Iran

    Razieh Jaberi

Authors
  1. Razieh Jaberi
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  2. Sara Mirsadeghi
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Contributions

RJ: performed cell culture, immunostainings, and other characterization assays, figure preparation, statistical analysis and wrote the paper, SM: contributed in writing and professional editing of the manuscript, SK: conceived the project, supervised the experiment and assessments, discovered the grant sources and also contributed in manuscript editing.

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Correspondence to Sahar Kiani.

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Jaberi, R., Mirsadeghi, S. & Kiani, S. In vitro characterization of subventricular zone isolated neural stem cells, from adult monkey and rat brain. Mol Biol Rep 48, 1311–1321 (2021). https://doi.org/10.1007/s11033-021-06201-7

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  • DOI: https://doi.org/10.1007/s11033-021-06201-7

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