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In vitro neural differentiation of CD34 + stem cell populations in hair follicles by three different neural induction protocols

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Abstract

Differentiation of hair follicle stem cells (HFSCs) into neurons and glial cells represents a promising cell-based therapy for neurodegenerative diseases. The hair follicle bulge area is reported as a putative source of new stem cell population for many years. In vitro studies have implicated neural differentiation of HFSCs. Here, we report the identification and purification of CD34 + cells from hair follicle by magnetic activated cell sorting (MACS). We next determined the cytotoxic effects of all-trans retinoic acid (RA) by using cell viability assays. Moreover, the neural differentiation potential of CD34 + cells was evaluated in the presence of RA, serum-free condition, and neural differentiation medium (NDM) treatments by using immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). Our results showed that the isolated CD34 + stem cells were 12% of the total cells in the bulge area, and the neural cells derived from the stem cells expressed nestin, microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein (GFAP). Interestingly, all the neural induction media supported neuronal differentiation most effectively, but treatment with serum-free medium significantly increased the number of GFAP-positive glial cells. Moreover, increasing RA concentration (≥10 μM) leads to increased cell death in the cells, but a lower concentration of RA (1 μM) treatment results in a decrease in CD34-expressing stem cells. These findings show an instructive neuronal effect of three neural induction media in HFSCs, indicating the important role of this induction media in the specification of the stem cells toward a neural phenotype.

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Acknowledgments

This research was supported by a grant from Ardabil University of Medical Sciences (Ardabil, Iran, No. 89350). The authors also wish to thank Dr Shahab Bohlooli for providing useful equipment.

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this article.

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Authors and Affiliations

  1. Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences and Pathology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Nowruz Najafzadeh, Mohsen Sagha, Mohammad Ghasem Golmohammadi, Nasim Massahi Oskoui & Maryam Deldadeh Moghaddam

  2. Department of Biological Sciences, Faculty of Basic Sciences, Urmia University, Urmia, Iran

    Shirin Heydari Tajaddod

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  1. Nowruz Najafzadeh
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  2. Mohsen Sagha
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  3. Shirin Heydari Tajaddod
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Correspondence to Nowruz Najafzadeh.

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Najafzadeh, N., Sagha, M., Heydari Tajaddod, S. et al. In vitro neural differentiation of CD34 + stem cell populations in hair follicles by three different neural induction protocols. In Vitro Cell.Dev.Biol.-Animal 51, 192–203 (2015). https://doi.org/10.1007/s11626-014-9818-2

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