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Microbiota of human inferior turbinate

ABSTRACT: Analysis of human inferior turbinate microbiota

PROVIDER: PRJEB14731 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1520641.fastq.gz	Fastqsanger.gz
	ERR1520642.fastq.gz	Fastqsanger.gz

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Project description:Adult human neural crest-derived stem cells are of extraordinary high plasticity and promising candidates for the use in regenerative medicine. Here we describe for the first time a novel neural crest-derived stem cell population within the respiratory epithelium of adult human inferior turbinate. In contrast to superior and middle turbinates, high amounts of source material could be isolated from human inferior turbinates using minimally-invasive surgery methods isolation is efficient even from older patients. Within their endogenous niche, inferior turbinate stem cells (ITSCs) expressed high levels of nestin, p75NTR and S100. Immuno-electron microscopy using anti-p75 antibodies displayed that ITSCs are of glial origin and closely related to non-myelinating Schwann cells. Cultivated ITSCs were positive for nestin and S100 as well as the neural crest markers Slug and Sox10. Whole genome microarray analysis showed pronounced differences to human ES cells in respect to pluripotency markers OCT4, SOX2, LIN28 and NANOG, whereas expression of WDR5, KLF4 and c-MYC was nearly similar. ITSCs were able to differentiate into ectodermal, mesodermal and endodermal cell types. Additionally ITSCs are able to survive and perform neural crest typical chain migration in vivo when transplanted into chicken embryos. However ITSCs do not form teratomas in immunodeficient mice. Finally, we developed a separation strategy based on magnetic cell sorting of p75 positive ITSCs, which formed larger neurospheres and proliferated faster than p75 negative ITSCs. Taken together our study describes a novel, readily accessible source of multipotent human neural crest-derived stem cells for potential cell-replacement therapy. RNA samples to be analyzed by microarrays were prepared using Qiagen RNeasy columns with on-column DNA digestion. 500 ng of total RNA per sample was used as input into a linear amplification protocol (Ambion) which involved synthesis of T7-linked double-stranded cDNA and 12 hours of in vitro transcription incorporating biotin-labelled nucleotides. Purified and labeled cRNA was then hybridized for 18h onto HumanHT-12 v4 expression BeadChips (Illumina) following the manufacturers instructions. After washing as recommended, chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using the iScan reader (Illumina) and accompanying software.

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Microbiota of human inferior turbinate

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