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

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

PROVIDER: PRJNA816483 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR18332087.fastq.gz	Fastqsanger.gz
	SRR18332088.fastq.gz	Fastqsanger.gz
	SRR18332089.fastq.gz	Fastqsanger.gz
	SRR18332090.fastq.gz	Fastqsanger.gz
	SRR18332091.fastq.gz	Fastqsanger.gz

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Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures

Project description:Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures

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

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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

Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures.

Project description:Adult autologous human epidermal stem cells can be extensively expanded ex vivo for cell and gene therapy. Identifying the mechanisms involved in stem cell maintenance and defining culture conditions to maintain stemness is critical, because an inadequate environment can result in the rapid conversion of stem cells into progenitors/transient amplifying cells (clonal conversion), with deleterious consequences on the quality of the transplants and their ability to engraft. Here, we demonstrate that cultured human epidermal stem cells respond to a small drop in temperature through thermoTRP channels via mTOR signaling. Exposure of cells to rapamycin or a small drop in temperature induces the nuclear translocation of mTOR with an impact on gene expression. We also demonstrate by single-cell analysis that long-term inhibition of mTORC1 reduces clonal conversion and favors the maintenance of stemness. Taken together, our results demonstrate that human keratinocyte stem cells can adapt to environmental changes (e.g., small variations in temperature) through mTOR signaling and constant inhibition of mTORC1 favors stem cell maintenance, a finding of high importance for regenerative medicine applications.

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Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures

Project description:Adult autologous human epidermal stem cells can be extensively expanded ex vivo for cell and gene therapy. Identifying the mechanisms involved in stem cell maintenance and defining culture conditions to maintain stemness is critical, because an inadequate environment can result in the rapid conversion of stem cells into progenitors/transient amplifying cells (clonal conversion), with deleterious consequences on the quality of the transplants and their ability to engraft. Here, we demonstrate that cultured human epidermal stem cells respond to a small drop in temperature through thermoTRP channels via mTOR signaling. Exposure of cells to rapamycin or a small drop in temperature induce the nuclear translocation of mTOR with an impact on gene expression. We also demonstrate by single-cell analysis that long-term inhibition of mTORC1 reduces clonal conversion and favors the maintenance of stemness. Taken together, our results demonstrate that human keratinocyte stem cells can adapt to environmental changes (e.g. small variations in temperature) through mTOR signaling and constant inhibition of mTORC1 favors stem cell maintenance, a finding of high importance for regenerative medicine applications.

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Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures

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2023-05-29 | GSE199523 | GEO

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