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

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.

Generating inner ear organoids containing putative cochlear hair cells from human pluripotent stem cells

“…This is consistent with the functional maturation of cochlear hair cells and the development of hearing in neonatal mice 54 , however to improve the utility of our inner ear organoid culture to the point where we can investigate the possible contribution of acoustic stimulation, we need to manipulate additional signalling molecules that could amplify the otic induction process. This could focus on various FGFs or retinoic acid (known to contribute to the control of location-specific aspects of hair cell phenotypes) 55 , 56 but a more tractable approach might be to extend the development of fluorescent reporter systems analogous to the ATOH-GFP cell line developed by Koehler et al 19…”

miR‐124 promotes neural differentiation in mouse bulge stem cells by repressing Ptbp1 and Sox9

“…overexpress during neural differentiation and enhance cell cycle exit by inhibiting cyclin D1, BAF53A, Ptbp1, and Sox9(Shenoy & Blelloch, 2014b). These results suggest that miR-124 and let-7 regulate adult neurogenesis in the adult brain as well as during embryogenesis.Different types of stem cells with varying differentiation potentialssuch as epidermal-neural crest stem cells (Epi-NCSCs;Hu, Zhang, & Sieber-Blum, 2006;Yu, Kumar, Kossenkov, Showe, & Xu, 2010), nestinpositive(Amoh, Li, Katsuoka, Penman, & Hoffman, 2005b), and CD34positive cells(Najafzadeh et al, 2015c) have been demonstrated in hair follicles. HFSCs can differentiate into neurons, astrocytes, oligodendrocytes, and Schwann cells.…”

Human nail stem cells are retained but hypofunctional during aging

“…CD29 is the most widely accepted of the skin stem cell markers identified (Bose et al 2013 ; Watt 1998 ). CD34 + cells are a rich population of skin epithelial stem cells (Hong et al 2014 ; Lin et al 2015 ; Najafzadeh et al 2015 ; Ouji et al 2015 ; Zhu et al 2013 ). The expression of Lgr6 has been observed in several adult skin stem cells (Barker et al 2013 ; Lehoczky and Tabin 2015 ; Snippert et al 2010 ).…”

“…We subsequently identified Lgr6 expression in K15 + cells. CD34 and CD29 are generally recognised as commonly occurring in skin epithelial stem cells and are widely accepted as stem cell markers (Bose et al 2013 ; Hong et al 2014 ; Najafzadeh et al 2015 ; Ouji et al 2015 ; Watt 1998 ; Zhu et al 2013 ). Additionally, their expression has been discovered in the same cells in human nails (Fig.…”