Isolated of dermal papilla cells from follicular unit extraction
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE142095 | GEO | 2022/12/01
REPOSITORIES: GEO
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Project description:Dermal papilla cells (DPCs) are located at the bottom of hair follicles and play important roles on hair induction by interacting with epidermal cells. DPCs are promising cell sources for hair regeneration therapy for alopecia patients. However, freshly isolated DPCs rapidly lose their hair inductive activity and proliferative potential under culture conditions. We examined the effects of telomere reverse transcriptase (TERT) and B-cell-specific Moloney murine leukemia virus insertion region 1 (BMI1) on extending the life span of murine DPCs as well as on hair inductive activity.TERT and BMI1 genes were introduced into murine DPCs using lentivirus vectors. Hair inductive activity of transfected DPCs (t-DPCs) was determined by in vivo chamber assay in nude mice. Transcripts between intact dermal papillae (DPs) and cultured DPCs were compared by microarray analysis.
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Project description:Dermal papilla cells (DPCs) and epidermal hair matrix cells (HMCs) located in hair bulb are the key cell types during the hair follicles (HFs) development. To explore the mRNA and miRNA expression of DPCs and HMCs of yak hair follicle, illustrating the mocular basis of the interaction and cellular communication between DPCs and HMCs during the hair follicle development, the DPCs and HMCs of yak were isolated and cultured, RNA-seq was used to identify the differentially expressed mRNAs and miRNAs between DPCs and HMCs.
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Project description:Dermal papilla cells (DPCs) and epidermal hair matrix cells (HMCs) located in hair bulb are the key cell types during the hair follicles (HFs) development. To explore the mRNA and miRNA expression of DPCs and HMCs of yak hair follicle, illustrating the mocular basis of the interaction and cellular communication between DPCs and HMCs during the hair follicle development, the DPCs and HMCs of yak were isolated and cultured, RNA-seq was used to identify the differentially expressed mRNAs and miRNAs between DPCs and HMCs.
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Project description:We investigated the effect of low oxygen culture on the proliferation and hair inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were cultured in atmospheric/hyperoxia (20% O2), physiological/normoxia (6% O2), or hypoxia (1% O2) conditions, respectively. Proliferation of DPCs and DSCs was highest under normoxia. Hypoxia inhibited proliferation of DPCs but enhanced proliferation of DSCs. In DPCs, hypoxia down-regulated expression of hair inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN, and normoxia up-regulated expression of ALP. In DSCs, both normoxia and hypoxia up-regulated SOX2 expression, and hypoxia down-regulated BMP4 expression. Microarray analysis revealed increased expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, under hypoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In DPCs, normoxia allowed the most efficient induction of hair follicles. In DSCs, hypoxia allowed the most efficient induction and maturation of hair follicles. These results suggest that low oxygen culture enhances the proliferation and maintains functions of human DPCs and DSCs and could be used for skin engineering and clinical applications.
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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.
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Project description:Human scalp hair follicles were snapfrozen directly after surgery or organ-cultured in the presence/absence of SEP1 phages. After treatment total RNA was isolated and prepared for RNA-seq
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Project description:We investigated the effect of low oxygen culture on the proliferation and hair inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were cultured in atmospheric/hyperoxia (20% O2), physiological/normoxia (6% O2), or hypoxia (1% O2) conditions, respectively. Proliferation of DPCs and DSCs was highest under normoxia. Hypoxia inhibited proliferation of DPCs but enhanced proliferation of DSCs. In DPCs, hypoxia down-regulated expression of hair inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN, and normoxia up-regulated expression of ALP. In DSCs, both normoxia and hypoxia up-regulated SOX2 expression, and hypoxia down-regulated BMP4 expression. Microarray analysis revealed increased expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, under hypoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In DPCs, normoxia allowed the most efficient induction of hair follicles. In DSCs, hypoxia allowed the most efficient induction and maturation of hair follicles. These results suggest that low oxygen culture enhances the proliferation and maintains functions of human DPCs and DSCs and could be used for skin engineering and clinical applications.
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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),
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Project description:Alopecia is an exceedingly prevalent problem and lacks effective therapy. Recently, research has focused on early-passage dermal papilla cells (DPCs), which have hair inducing activity both in vivo and in vitro. Our previous study indicated that factors secreted from early-passage DPCs contribute to hair follicle (HF) regeneration. To identify which factors are responsible for HF regeneration and why late-passage DPCs lose this potential，we collected 48-h-culture medium (CM) from both of passage 3 and 9 DPCs and subcutaneously injected the DPC-CM into NU/NU mice. Passage 3 DPC-CM induced HF regeneration, based on the emergence of a white hair coat, but passage 9 DPC-CM not. In order to identify the key factors responsible for hair inductive capacity, CM from passage 3 and 9 DPCs was analyzed by iTRAQ-based quantitative proteomic technology. We identified 1360 proteins, of which 213 proteins were differentially expressed between CM from early-passage vs. late-passage DPCs, including SDF1, MMP3, biglycan and LTBP1.Further analysis indicated that the differentially-expressed proteins regulated the Wnt, TGF-β and BMP signaling pathways, which directly and indirectly participate in HF morphogenesis and regeneration. Subsequently, we selected 19 proteins for further verification by multiple reaction monitoring (MRM) between the two types of CM. These results would be used to identify the key factors for inducing HF regeneration and reveal the molecular mechanisms of losing inductive ability of DPCs. Furthermore, it is possible to explore some drugs for alopecia in the clinic according to this secretome date analysis.
2018-10-26 | PXD004201 | Pride
Project description:Hair follicle growth inhibitory effect of dihydrotestosterone (DHT)-treated two-dimensional (2D)- and 3D-cultured DPCs were evaluated. 2D- and 3D-cultured DPC proliferation was inhibited after co-culturing with outer root sheath (ORS) cells under DHT treatment. Moreover, gene expression levels of β‐catenin and neural cell adhesion molecule (NCAM) were significantly decreased and those of cleaved caspase-3 significantly increased in 2D- and 3D-cultured DPCs with increasing DHT concentrations. ORS cell proliferation also significantly increased after co-culturing in the control-3D model compared with the control-2D model. ki67 downregulation and cleaved caspase-3 upregulation in DHT-2D and 3D groups significantly inhibited ORS cell proliferation. Sequencing showed an increase in the expression of genes related to extracellular matrix synthesis in the 3D model group. Additionally, the top 10 hub genes were identified, and the expression of nine chemokine-related genes in DHT-treated DPCs was found significantly increased. We also identified the interactions between transcription factor (TF)–genes and microRNAs with hub genes and the TF–microRNA coregulatory network. Overall, the findings indicate that 3D-cultured DPCs are more representative of in vivo conditions than are 2D-cultured DPCs, and contribute to our understanding of the molecular mechanisms underlying androgen-induced alopecia.
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