Project description:The dermal sheath cup is the peribulbar component of the hair follicle dermal sheath, and has hair inductive potential similar to the dermal papilla. To characterize it in comparison with other mesenchymal follicle tissuesparts, we performed gene expression profiling of intact dermal sheath cups, which were separated from hair follicles by microdissection. Gene expression profiles of the dermal sheath cup, dermal papilla and upper dermal sheath were compared. We identified a dermal sheath cup signature composed of 32 upregulated genes, which included extracellular matrix components and BMP binding mollecules, while dermal papilla signature included a number of dermal papilla signature genes which had already reported. Analyses of upstream regulators showed that TGF- b1 is a putative regulator of these genes. These results suggest some of molecular mechanism that contributes to human dermal sheath cup properties, which could be useful for hair follicle bioengineering.
Project description:Here we have developed a novel FACS strategy to prospectively isolate hair follicle dermal stem cells, dermal sheath and dermal papilla cells from adult skin initiating synchronous hair follicle regeneration and identified Hic1 as a marker of hfDSCs and Rspondins as stimulators of hfDSCs and epithelial cells, and subsequently hair follicle regeneration
Project description:While this is available for murine HFs, systematic characterization of the transcriptome of specific human hair follicle (HF) compartments remains a major unmet challenge in human hair research. Here, we have aimed to initiate the generation of such an atlas to reveal HF compartment-specific transcriptome. We performed laser capture microdissection (LCM) on 8 selected compartments, i.e. bulge outer root sheath (ORS), suprabulbar ORS, inner root sheath (IRS), precortical hair matrix (HM), germinative HM, dermal papilla (DP), the lower dermal cup (DC), and the connective tissue sheath (CTS), of human anagen VI scalp HFs, and analysed these by RNA-seq. Principal component analysis (PCA) was used to validate the distinctness of the microdissected compartments. Biological replicates were clearly separated by HF compartments and a high degree of similarity of the distinct transcriptional profiles of each examined compartment was seen. Afterwards, we matched well-known signature genes to the correct HF compartments. For example, VCAN (versican) and ALPL (alkaline phosphatase) were correctly expressed in the DP, the lower DC, and suprabulbar CTS, while TCHH (trichohyalin) was solely found in the proximal IRS, and KRT85 (Keratin 85) and KRT31 (Keratin 31) were almost exclusively expressed in the precortical HM. These transcriptomic data were used to generate an easily accessible database for identifying and defining compartment-specific gene expression profiles, which can be ultimately made publicly accessible (“hair follicle MONasterium InteraCtive atlas – hairMONic”). This human HF transcriptomic map and the associated interactive database will support the identification of HF compartment-specific signature genes, and facilitates the development of compartment-specific, therapeutic interventions for the management of human HF disorders.
Project description:Dermal papilla cells isolated from the human hair follicle are capable of inducing hair growth in recipient epithelia. However, demonstrating disparity from rodent dermal papilla, human cells lose this inductive competance immediately upon growth in culture under normal growth conditions. We grew dermal papilla cells in hanging drop cultures that are morphologically akin to intact dermal papilla, and found that by enhancing the environment for aggregation, we could restore the inductive capacity of human dermal papilla cells in culture. The underlying genes that regulate the inductive potential of dermal papilla cells is not well understood, and we sought to use global profiling to identify key genes and pathways related to inductive competance within dermal papilla cells. We used Affymetrix microarrays to profile human dermal papilla cells in both hair inducing, and non-hair inducing states.
Project description:Dermal papilla cells isolated from the human hair follicle are capable of inducing hair growth in recipient epithelia. However, demonstrating disparity from rodent dermal papilla, human cells lose this inductive competance immediately upon growth in culture under normal growth conditions. We grew dermal papilla cells in hanging drop cultures that are morphologically akin to intact dermal papilla, and found that by enhancing the environment for aggregation, we could restore the inductive capacity of human dermal papilla cells in culture. The underlying genes that regulate the inductive potential of dermal papilla cells is not well understood, and we sought to use global profiling to identify key genes and pathways related to inductive competance within dermal papilla cells. We used Affymetrix microarrays to profile human dermal papilla cells in both hair inducing, and non-hair inducing states. Affymetrix microarrays were used to to perform profiling of human dermal papilla cells, both as intact tissues (freshly isolated from scalp), and at several stages in subsequent two dimensional culture; cell explant outgrowths (p0), cells at passage 1 (p1), passage 3 (p3) and passage 5 (p5). RNA was isolated from cultured cells 72 hours after feeding. Cells at passage 3 were also grown in hanging drops to form dermal spheroids, that were used for RNA collection 48 hours after establishment. All experiments were performed using tissue from three biological replicates (#D5, D6, D7),
Project description:Our data showed that miR-221 had a profound effect on the biological function of dermal papilla cells (DPCs) and dermal sheath cells (DSCs) of hair follicle. Overexpression of miR-221 affected the MAPK signaling in DPCs, and PI3K/AKT signaling in DSCs.
Project description:We investigated epidermis derived fibroblasts in neonatal skin at single cell-transcriptomic levels. We found that keratin 5 lineage mesenchymal cells mainly showed the transcriptomic signatures as dermal papilla and dermal sheath cells. Our data suggested that hair follicle dermal cells were originally derived from epidermis via epithelial to mesenchymal transition.
Project description:RNA-Seq of human fetal and adult scalp hair follicle dermal papilla (DP), dermal sheath cup (DSC), and interfollicular dermal (IFD) cells
Project description:Much is still unknown about the molecular regulatory networks which govern the dermal papilla’s (DP) ability to induce hair follicle regeneration, a capacity which gradually decreases with age. DP and dermal sheath cup (DSC) cells from mature, anagen phase, hair follicles were manually microdissected from fresh frozen sections of 16-18 week human fetal scalp and 30-60 year old adult male scalp. Interfollicular dermal (IFD) fibroblasts were harvested for comparison. RNA-seq libraries from each cell population were prepared with Nugen’s Ovation RNA-Seq and Ultralow Library systems, sequenced to approximately 100 million total reads. A set of 121 genes was identified as significantly upregulated in fetal DP cells, as compared to both fetal DSC and IFD populations. Wnt/β-catenin, Shh, FGF, BMP, and Notch signaling pathways were significantly enriched among those genes differentially expressed between fetal and adult DP cells. Among them, Spondin-1, a Wnt agonist, was chosen for verification and can rescue hair follicle regeneration in skin reconstitution assays using cells from adult mice. Additionally, twenty-nine transcription factors were significantly upregulated in fetal DP compared to adult DP cells. Of those, seven transcription factor binding motifs were significantly enriched in the candidate promoter regions of differentially expressed genes between fetal and adult DP cells, suggesting a combinatorial regulatory role in the fetal DP phenotype. Further investigation into these regulators is warranted to determine if these proteins and/or associated pathways plays a role in the maintenance or modulation of DP cells to induce hair regeneration.