Project description:Clonogenic keratinocyte stem cells isolated from the bulge area of human telogen follicles were co-cultured with dermal papilla cells in a transwell system. RNA was isolated from stem cells for different periods of time (day 0, 1, 2, and 5) after co-culture with DP and analyzed for changes in gene expression using Genechip microarrays. Keywords = epithelial stem cells Keywords = DP Keywords = co-culture Keywords: other
Project description:Clonogenic keratinocyte stem cells isolated from the bulge area of human telogen follicles were co-cultured with dermal papilla cells in a transwell system. RNA was isolated from stem cells for different periods of time (day 0, 1, 2, and 5) after co-culture with DP and analyzed for changes in gene expression using Genechip microarrays.
Project description:During development, dermal papilla precursor cells (DPPCs) initiate embryonic hair follicle (HF) formation with epidermal placode cells. Obtaining DPPCs with trichogenic ability is critical for human HF regeneration because dermal papilla cells (DPCs) rapidly loseDuring development, dermal papilla precursor cells (DPPCs) initiate embryonic hair follicle (HF) formation with epidermal placode cells. Obtaining DPPCs with trichogenic ability is critical for human HF regeneration because dermal papilla cells (DPCs) rapidly lose their trichogenic ability in culture. Here, we generated trichogenic DPPCs from human induced pluripotent stem cells (iPSCs) via neural crest stem cells (NCSCs), based on the developmental evidence at the hair placode stage. SDC1+CD133− cells showed signature DP gene expression, spontaneous sphere formation and represented intermediate population in the differentiation way from NCSCs to DPCs. hiPSC-derived DPPCs generated HF equivalents in vitro and reconstituted de novo human HFs in vivo combined with hiPSC-derived epithelial stem cells. Remarkably, trichogenic ability of DPPC was only proven in the specific time window, providing insights into the loss of trichogenicity in cultured DPC. Thus, this study provides an in vitro model for studying DPC development and biology.
Project description:The use of dermal papilla cells for hair follicle (HF) regeneration is long accepted much attention. However, cultured dermal papilla cells tend to lose the hair-inducible capability during passaging, which restricts its application. Increasing evidences indicate that dermal papilla cells exert their regulatory function of HF growth mainly through their unique paracrine properties, opening up a way to exosome therapies.This study aimed to explore the effects of exosomes from high and low-passaged human scalp follicle dermal papilla cells (DP-Exos) on hair follicle stem cells (HFSCs) activation and hair growth, and to investigate the underline mechanism. DP-Exos were isolated by ultracentrifugation and cultured with human scalp follicles and HFSCs. The hair elongation and cell proliferation was assessed. Quantitative real-time PCR (qRT-PCR) and Western-blot were performed to detect the expression levels of a class of miRNAs and proteins which have positive roles in regulating hair growth and HFSCs proliferation. High throughput miRNA sequencing of miRNAs in high (P8) and low-passaged (P3) DP-Exos was performed, and the utmost miRNA and its target gene was identified via bioinformatics analysis.
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: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:Analysis of ex vivo isolated lymphatic endothelial cells from the dermis of patients to define type 2 diabetes-induced changes. Results preveal aberrant dermal lymphangiogenesis and provide insight into its role in the pathogenesis of persistent skin inflammation in type 2 diabetes. The ex vivo dLEC transcriptome reveals a dramatic influence of the T2D environment on multiple molecular and cellular processes, mirroring the phenotypic changes seen in T2D affected skin. The positively and negatively correlated dLEC transcripts directly cohere to prolonged inflammatory periods and reduced infectious resistance of patients´ skin. Further, lymphatic vessels might be involved in tissue remodeling processes during T2D induced skin alterations associated with impaired wound healing and altered dermal architecture. Hence, dermal lymphatic vessels might be directly associated with T2D disease promotion.
Project description:Analysis of ex vivo isolated lymphatic endothelial cells from the dermis of patients to define type 2 diabetes-induced changes. Results preveal aberrant dermal lymphangiogenesis and provide insight into its role in the pathogenesis of persistent skin inflammation in type 2 diabetes. The ex vivo dLEC transcriptome reveals a dramatic influence of the T2D environment on multiple molecular and cellular processes, mirroring the phenotypic changes seen in T2D affected skin. The positively and negatively correlated dLEC transcripts directly cohere to prolonged inflammatory periods and reduced infectious resistance of patients´ skin. Further, lymphatic vessels might be involved in tissue remodeling processes during T2D induced skin alterations associated with impaired wound healing and altered dermal architecture. Hence, dermal lymphatic vessels might be directly associated with T2D disease promotion. Global gene expression profile of normal dermal lymphatic endothelial cells (ndLECs) compared to dermal lymphatic endothelial cells derived from type 2 diabetic patients (dLECs).Quadruplicate biological samples were analyzed from human lymphatic endothelial cells (4 x diabetic; 4 x non-diabetic). subsets: 1 disease state set (dLECs), 1 control set (ndLECs)