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Maayke Kuijten, Claire Ginn, Peng Khaw, G Astrid Limb, Steve Brocchini; Identification and characterization of neural crest stem cells in rat olfactory mucosa as a use for cell-mediated neuroprotection as treatment of glaucoma. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1359.
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© ARVO (1962-2015); The Authors (2016-present)
The olfactory mucosa (OM) is an area of constant neural regeneration populated by olfactory ensheathing cells (OEC), which may constitute a source of neurotrophic support for spinal cord damage and glaucoma treatment. However, the heterogeneous nature of these cells makes their isolation difficult for therapeutic use. We examined the OM to identify neural crest stem cell (NCSC) populations, from which OECs originate and which may constitute an alternative source for neuroprotection in glaucoma.
We performed immunohistochemistry and confocal microscopy to characterise the rat OM. Different cell sorting strategies such as flow cytometry and differential adhesion were used to separate different cell populations. Analyses were performed using Western blotting and RT-PCR techniques.
Confocal analysis of postnatal day 16 OM tissue showed the presence of three different cell populations in the lamina propria (LP). Positive staining for mesenchymal stem cell (MSC) markers in this tissue indicates the presence of mesenchymal-like stem cells and their location seems to indicate a MSC niche in the LP of OM tissue. Expression of neural stem cell markers including Sox2, Pax6 and nestin was found in the LP, which could indicate the presence of different stem cell populations. The NCSC markers Sox10, snail and HNK-1 were also found, indicating the presence of a NCSC population in the LP. Differential adhesion has been used as a means of selecting different cell populations. In these cell populations, expression of the NCSC genes p75, Sox10, HNK-1 and snail was also seen. Expression of the above markers was further confirmed by examination of mRNA and protein lysates of whole OM. In addition, mRNA samples of the obtained cell population and OM tissue have been investigated for neurotrophin expression, such as BDNF, CNTF, GDNF, NGF, NT3 and NT4/5, which are all expressed.
The OM is a complex tissue that contains at least one or possibly more stem cell populations residing in the LP. Based on the gene expression profile, Western blotting of tissue and cell lysates and staining of tissue and cell populations, neural crest stem cells were shown to be present in the OM. In addition, the obtained cell populations and whole OM express several neurotrophin genes, suggesting their potential neuroprotective ability.
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