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Gene expression profiling of total RNA isolated from hiPSC-derived DPPCs, hiPSC-derived NCSCs, cultured hDPC spheres, cultured hDPC

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE100793 | GEO | 2023/06/25

REPOSITORIES: GEO

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Human dermal papilla cell culture medium iTRAQ

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

AR/miR-221/IGF-1 pathway mediates the pathogenesis of androgenetic alopecia

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Detection of the expression profiles of mRNA and miRNA between dermal papilla cells and hair matrix cells of yak hair follicle [RNA-seq]

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Introduction of the TERT and BMI1 Genes into Murine Dermal Papilla Cells Ameliorates Hair Inductive Activity

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: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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Transcription profiles of human dermal sheath cells cultured under 20%-O2 or 6%-O2 condition

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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Transcription profiles of human dermal papilla cells cultured under 20%-O2 or 6%-O2 condition

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Î²-cateinin activation in hair follicle dermal sheath induces ectopic hair outgrowth and skin fibrosis

Project description:Dermal sheath (DS) shows potent hair inducing capability and high plasticity without leading to immuno rejection. A recent study showed a subset of DS cells, identified as hair follicle (HF) dermal stem cells, can be mobilized to regenerate DS, maintain/supply the cell number of dermal papilla (DP) and modulate hair type. However, it is unclear how Wnt/Î²-catenin signaling regulates DS cells behaviors. Here we report that activation of Î²-catenin in DS, to some extent, endows it with hair inducing ability, reprogramming HF epidermal cells to generate new outgrowth. The new formed dermal condensates (DC)/DP lying adjacent the outgrowth derives from DS and/or its progeny, and homeostasis of pre-existing HFs is disturbed. Additionally, progressive skin fibrosis is prominent in hypodermis, where the excessive activated fibroblasts at least partially originate from DS and/or its progeny. Gene expression analysis of purified DS cells revealed that 44% DC signature genes are regulated, most of which are up-regulated. We found elevated expression of several growth factors, including Noggin, Fgf7 and Fgf10, which were previously implicated in HF induction. In summary, we confirm the high plasticity of DS cells by in vivo assays and report a mechanism by which Wnt/Î²-catenin signaling controls DS cells behaviors. We prospectively isolated control and cÎex3 DS cells (YFP+ve CD34-ve ALP-ve ITGÎ±8-ve cells) from P15 dorsal skins by FACS.

2016-05-31 | E-GEOD-81615 | biostudies-arrayexpress

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