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Comparative transcriptome profiling of human epidermis keratinocyte stem cells (KSC) versus keratinocyte progenitors (KP)

ABSTRACT: To characterize the global transcriptome of human keratinocyte stem cells (KSC) and keratinocyte progenitors (KP), primary basal keratinocyte subpopulations enriched in quiescent stem cells [Itg-α6bright / Trf-Rdim] or in cycling progenitors [Itg-α6bright / Trf-Rbright] were purified by flow cytometry from human skin samples. Sorted keratinocytes were then seeded in culture plates and maintained in culture medium overnight (15 hours), and genome-wide transcriptome analysis of these two subpopulations was directly performed, in order to globally maintain the initial phenotypes. After RNA preparation, gene profiling was performed using oligonucleotide microarrays (26068 probes). LOWESS normalisation was applied. Keywords: epidermis, keratinocyte stem cells, progenitors, transcriptome, stemness

ORGANISM(S): Homo sapiens

PROVIDER: GSE68583 | GEO | 2015/05/06

SECONDARY ACCESSION(S): PRJNA283140

REPOSITORIES: GEO

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Comparative transcriptome profiling of human epidermis keratinocyte stem cells (KSC) versus keratinocyte progenitors (KP)

Project description:To characterize the global transcriptome of human keratinocyte stem cells (KSC) and keratinocyte progenitors (KP), primary basal keratinocyte subpopulations enriched in quiescent stem cells [Itg-?6bright / Trf-Rdim] or in cycling progenitors [Itg-?6bright / Trf-Rbright] were purified by flow cytometry from human skin samples. Sorted keratinocytes were then seeded in culture plates and maintained in culture medium overnight (15 hours), and genome-wide transcriptome analysis of these two subpopulations was directly performed, in order to globally maintain the initial phenotypes. After RNA preparation, gene profiling was performed using oligonucleotide microarrays (26068 probes). LOWESS normalisation was applied. Keywords: epidermis, keratinocyte stem cells, progenitors, transcriptome, stemness Dye-swap hybridization were performed. Slides were scanned with a Genepix 4000 microarray scanner (Axon Instruments, Molecular devices, Sunnyvale, CA). For each hybridized spot, the Cy3 and Cy5 fluorescence values were obtained by using Genepix Pro 4.0 software (Axon Instruments) and were saved as a result file

2015-05-06 | E-GEOD-68583 | biostudies-arrayexpress

Homo sapiens

Project description:Comparative transcriptome profiling of human epidermis keratinocyte stem cells (KSC) versus keratinocyte progenitors (KP)

| PRJNA283140 | ENA

Transcriptome of irradiated keratinocyte stem cells

2007-05-05 | GSE7693 | GEO

Short-Term Treatment with Rho-Associated Kinase Inhibitor Preserves Keratinocyte Stem Cell Characteristics In Vitro

Project description:Primary keratinocytes including keratinocyte stem cells (KSCs) can be cultured as epidermal sheets in vitro and are attractive for cell and gene therapies for genetic skin disorders. However, the initial slow growth of freshly isolated keratinocytes hinders clinical applications. Rho-associated kinase inhibitor (ROCKi) has been used to overcome this obstacle, but its influence on the characteristics of KSC and its safety for clinical application remains unknown. In this study, primary keratinocytes were treated with ROCKi Y-27632 for six days (short-term). Significant increases in colony formation and cell proliferation during the six-day ROCKi treatment were observed and confirmed by related protein markers and single-cell transcriptomic analysis. In addition, short-term ROCKi-treated cells maintained their differentiation ability as examined by 3D-organotypic culture. However, these changes could be reversed and became indistinguishable between treated and untreated cells once ROCKi treatment was withdrawn. Further, the short-term ROCKi treatment did not reduce the number of KSCs. In addition, AKT and ERK pathways were rapidly activated upon ROCKi treatment. In conclusion, short-term ROCKi treatment can transiently and reversibly accelerate initial primary keratinocyte expansion while preserving the holoclone-forming cell population (KSCs), providing a safe avenue for clinical applications.

2023-01-17 | GSE207130 | GEO

Amniotic Fluid primary human keratinocyte culture

Project description:Amniotic Fluid primary human keratinocyte culture

| PRJNA757450 | ENA

The impact of HPV8 E6 on Lrig1+ hair follicle stem cell populations

Project description:Despite epidermal turnover, the skin is host to a complex array of microbes including viruses, such as the human papillomavirus (HPV), which must infect and manipulate skin keratinocyte stem cells (KSC) to survive. This crosstalk between the virome and KSC populations remains largely unknown. Here, we investigated the effect of HPV8 on KSCs using various mouse models. We observed that the HPV8 early region gene E6 specifically caused Lrig1+hair follicle junctional zone KSC proliferation and expansion, which would facilitate viral transmission. Within Lrig1+KSCs specifically, HPV8 E6 bound intracellular p300 to phosphorylate the STAT3 transcriptional regulatory node. This induces a transcriptional switch from TAp63 to ΔNp63 expression, resulting in expansion of KSC into the overlying epidermis. HPV8 was associated with 70% of human actinic keratoses (AK). Together these results redefine human AK as an expansion of KSC, which lack protecting melanosomes and are thus susceptible to sun-light-induced malignant transformation.

2024-02-22 | GSE247466 | GEO

Transcriptome of irradiated keratinocyte stem cells

Project description:To characterise the response of keratinocyte stem cells to ionizing radiation, gene profiling was performed using oligonucleotide microarrays (26068 probes). Cells were seeded in culture plates after cell sorting, irradiated the next day (2 Gy at 15 hours in culture), and RNA was prepared 3 hours after exposure. LOWESS normalisation was applied. dye-swap hybridization were performed. Slides were scanned with a Genepix 4000 microarray scanner (Axon Instruments, Molecular devices, Sunnyvale, CA). For each hybridized spot, the Cy3 and Cy5 fluorescence values were obtained by using Genepix Pro 4.0 software (Axon Instruments) and were saved as a result file

2010-06-11 | E-GEOD-7693 | biostudies-arrayexpress

Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures [RNA-seq]

Project description:Purpose: Comparison of genes expression pattern in normal human epidermal keratinocytes (NHEKs) which were cultured at several specific temperatures or with rapamycin. Methods: Basically, NHEKs were cultured with irradiated 3T3J2 feeder cells in cFAD medium (Pr Green's method) for 1 week. NHEKs were cultured at 32, 35, 36, 37 and 38 degrees C from beginning to end of cell culture, respectively. Furthermore, 100 nM rapamycin-added cFAD medium and ed at 37 degrees C. 7days after cell cuture, NHEKs were harvested for RNAseq analysis. Results: The culture condition at 32 degrees C and 37 degrees C with rapamycin maintained human epidermal keratinocyte stem cells. Conclusions: mTORC1 inhibition via temperature changes favors ex vivo maintenance of human epidermal keratinocyte stem cells.

2023-05-29 | GSE199522 | GEO

Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures [miRNA-seq]

2023-05-29 | GSE198696 | GEO

MicroRNA-198 an inhibitor of keratinocyte migration

Project description:This study identifies miR-198 as a potential inhibitor of keratinocyte migration in skin This study identifies genes differentially expressed during the early phases of wound healing using an ex vivo organ culture model of human skin

2013-03-02 | GSE37967 | GEO

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