Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human dermal pericytes compared to the remaining dermal cells of neonatal human foreskin

ABSTRACT: Pericytes derived from skin dermis can substantially enhance the short-term tissue-regenerative capacity of human epidermal cells already committed to differentiation; they also display both phenotypic and functional properties of mesenchymal stem cells. In this microarray analysis, we compared the gene expression profile of dermal pericytes to that of the remaining dermal cells of neonatal human foreskin. Experiment Overall Design: Human neonatal foreskin was digested overnight in dispase II at 4Â°C to separate the epidermis from the dermis. Subsequently the dermis was digested for 1-2 hours at 37Â°C in a mixed dispase and collagenase solution and then fractionated into two populations, i.e. pericytes (HD-1bri) and the remaining dermal cells (HD-1dim), on the basis of differential VLA-1 expression using fluorescence-activated cell sorting. Total RNA from 15,000 cells of each population was extracted from 4 independent replicate sorts. mRNAs were amplified using a T7-primer-based-2-round linear RNA amplification protocol (GeneChip Two-Cycle cDNA synthesis kit). Fragmented and biotin-labelled cRNA from each individual sample was hybridised to Affymetrix HG-U133 plus 2.0 arrays and scanned on a Affymetrix GeneChip scanner. Probe intensities were RMA normalized and log2-transformed expression values were compared using moderated t statistics to quantify differences between individual samples.

ORGANISM(S): Homo sapiens

SUBMITTER: Holger Schlueter

PROVIDER: E-GEOD-17014 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A role for pericytes as microenvironmental regulators of human skin tissue regeneration.

Paquet-Fifield Sophie S Schlüter Holger H Li Amy A Aitken Tara T Gangatirkar Pradnya P Blashki Daniel D Koelmeyer Rachel R Pouliot Normand N Palatsides Manuela M Ellis Sarah S Brouard Nathalie N Zannettino Andrew A Saunders Nick N Thompson Natalie N Li Jason J Kaur Pritinder P

The Journal of clinical investigation 20090803 9

The cellular and molecular microenvironment of epithelial stem and progenitor cells is poorly characterized despite well-documented roles in homeostatic tissue renewal, wound healing, and cancer progression. Here, we demonstrate that, in organotypic cocultures, dermal pericytes substantially enhanced the intrinsically low tissue-regenerative capacity of human epidermal cells that have committed to differentiate and that this enhancement was independent of angiogenesis. We used microarray analysi ...[more]

PMID: 19652362

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Project description:Hair follicle formation depends on reciprocal epidermal-dermal interactions and occurs during skin development, but not in adult life. This suggests that the properties of dermal fibroblasts change during postnatal development. To examine this, we used a PdgfraEGFP mouse line to isolate GFP-positive fibroblasts from neonatal skin, adult telogen and anagen skin and adult skin in which ectopic hair follicles had been induced (EF skin) by transgenic epidermal activation of beta-catenin. We also isolated epidermal cells from each mouse. The gene expression profile of EF epidermis was most similar to that of anagen epidermis, consistent with activation of beta-catenin signalling. In contrast, adult dermis with ectopic hair follicles more closely resembled neonatal dermis than adult telogen or anagen dermis. In particular, genes associated with mitosis were upregulated and extracellular matrix-associated genes were downregulated in neonatal and EF fibroblasts. We confirmed that sustained epidermal beta-catenin activation stimulated fibroblasts to proliferate to reach the high cell density of neonatal skin. In addition, the extracellular matrix was comprehensively remodelled, with mature collagen being replaced by collagen subtypes normally present only in developing skin. The changes in proliferation and extracellular matrix composition originated from a specific subpopulation of fibroblasts located beneath the sebaceous gland. Our results show that adult dermis is an unexpectedly plastic tissue that can be reprogrammed to acquire the molecular, cellular and structural characteristics of neonatal dermis in response to cues from the overlying epidermis. We have isolated the following populations of cells from mouse back skin by flow cytometry: 1A) GFP+ WT neonatal dermal fibroblasts, 1B) ItgA6+ WT neonatal epidermal keratinocytes, 2A) GFP+ WT telogen dermal fibroblasts, 2B) ItgA6+ WT telogen epidermal keratinocytes, 3A) GFP+ D2 transient activation (anagen) dermal fibroblasts, 3B) ItgA6+ D2 transient activation (anagen) epidermal keratinocytes, 4A) GFP+ D2 sustained activation (ectopic follicles) dermal fibroblasts, 4B) ItgA6+ D2 sustained activation (ectopic follicles) epidermal keratinocytes

Dataset Information

Transcription profiling of human dermal pericytes compared to the remaining dermal cells of neonatal human foreskin

Publications

A role for pericytes as microenvironmental regulators of human skin tissue regeneration.

Similar Datasets

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