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: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:Transcriptome analysis of hTERT-immortalized balding (BAB) and non-balding (BAN) dermal papilla cells derived from frontal and occipital scalp of male patients with androgenetic alopecia Hamilton grade IV. Interrogation of transcriptome differences between BAB and BAN after dihydrotestosterone (DHT, active metabolite of androgen) treatment revealed significant enrichment of vasculature-related genes among down-regulated genes in BAB compared to BAN.
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: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: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 derived from 3 individual donors were propagated and cultured using the hanging drop method (Higgins et al., PNAS 2013). Cells were cultured in drugs for two days
Project description:<p>Background Androgenetic alopecia (AGA), the most prevalent form of hair loss, is driven by the dysfunction of dermal papilla cells (DPCs). Emerging evidence implicates DPC senescence in the pathogenesis of AGA; however, the underlying molecular mechanisms remain incompletely elucidated.</p><p>Methods We employed a multi-faceted experimental approach, including analyses of human scalp tissues, primary DPCs, a dihydrotestosterone (DHT)-induced AGA mouse model, and immortalized DPCs stimulated with DHT. Cellular senescence was evaluated via expression of senescence markers (p16INK4a, p21, p53) and senescence-associated β-galactosidase staining. Cell proliferation, migration, and apoptosis were also assessed. Mitochondrial function was evaluated using transmission electron microscopy, MitoTracker, MitoSOX, JC-1, and Seahorse assays. RNA sequencing and bioinformatics analyses were performed to identify differentially expressed genes. The interaction between Aconitate Decarboxylase 1 (ACOD1) and DDX1 was verified via co-immunoprecipitation, mass spectrometry, and molecular docking. Metabolomic analysis was performed to profile intracellular metabolic alterations. Functional experiments included ACOD1 knockdown/overexpression and exogenous supplementation with 4-octyl itaconate (4-OI). Hair follicle morphology and hair loss in AGA mice were evaluated following 4-OI treatment.</p><p>Results Senescence markers were significantly elevated in AGA DPCs, accompanied by increased senescence-associated β-galactosidase staining, reduced cell proliferation and migration, and enhanced apoptosis. DHT-induced mitochondrial dysfunction in DPCs was characterized by increased mitochondrial fragmentation, decreased mitochondrial cristae, superoxide accumulation, reduced membrane potential, and impaired oxidative phosphorylation. RNA sequencing identified ACOD1 as significantly downregulated in DHT-treated DPCs. ACOD1 knockdown induced mitochondrial dysfunction, cellular senescence, and functional impairment in DPCs, while ACOD1 overexpression rescued these DHT-induced phenotypes. Mechanistically, ACOD1 interacted with DDX1 to inhibit its methylation; ACOD1 knockdown enhanced DDX1 methylation, thereby promoting mitochondrial dysfunction and DPC senescence. Metabolomic analysis demonstrated that ACOD1 knockdown significantly reduced itaconate levels. Exogenous 4-OI supplementation reduced DDX1 methylation, ameliorated DHT-induced mitochondrial dysfunction and cellular senescence, promoted cell proliferation and migration, suppressed apoptosis, mitigated hair follicle miniaturization, and alleviated hair loss in AGA mice.</p><p>Conclusions Our findings reveal a novel mechanism underlying DPC senescence in AGA, wherein ACOD1 deficiency promotes DDX1 methylation, mitochondrial dysfunction, and subsequent DPC senescence. ACOD1 represents a promising therapeutic target for AGA, and 4-OI may have translational potential for the treatment of AGA.</p>
Project description:Our study developed a novel 3D co-culture system to mimic the in vivo ECM dynamics. Using this system, we examined the interaction between primary dermal papilla cells isolated from patients with neonatal keratinocytes.