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Single cell RNA-sequencing of EpCAM-, CD45-, CD31- NG2- murine mammary tumor fibroblasts

ABSTRACT: Single cell RNA-sequencing of EpCAM-, CD45-, CD31- NG2- murine mammary tumor fibroblasts

PROVIDER: PRJNA436229 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6790720.fastq.gz	Fastqsanger.gz
	SRR6790730.fastq.gz	Fastqsanger.gz
	SRR6790740.fastq.gz	Fastqsanger.gz
	SRR6790750.fastq.gz	Fastqsanger.gz
	SRR6790760.fastq.gz	Fastqsanger.gz

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Single cell RNA-sequencing of EpCAM-, CD45-, CD31- NG2- murine mammary tumor fibroblasts

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Project description:To further development of our gene expression approach to biodosimetry, we have employed microRNA microarray expression profiling to identify genes with the potential to distinguish liver metastasis related microRNA. Colorectal cancer patients were administered anesthesia and 20 mL BM was taken from the right and left anterior iliac crests before surgery. Mononucleated cells were collected using a standard Ficoll-Hypaque gradient technique. To enrich for EpCAM+ cells, CD14+ cells were removed from the whole bone marrow using auto MACSTM pro (Milteny Biotec, Bergisch Gladbach, Germany) with anti-CD14 immunomagnetic beads (clone; TÜK4, Milteny Biotec). Next, CD45+ cells were removed by treatment with anti-CD45 immunomagnetic beads (clone; 5B1; Milteny Biotec). The residual CD14−CD45− cells were then incubated with FcR blocking reagent (Milteny Biotec), followed by incubation with anti-EpCAM immunomagnetic beads (clone; HEA-125, Milteny Biotec), and the CD14−CD45−EpCAM+ cells were taken up. Total RNA of these cells we analyzed the microRNA levels of CD14−CD45−EpCAM+ cells obtained from non-metastasis patients (n = 12) and liver metastasis patients (n = 7). Ten-microRNA consensus signature was identified that distinguished between CD14−CD45−EpCAM+ cells from liver metastasis patients and CD14−CD45−EpCAM+ cells from non-liver metastasis patients.

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