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RNA-seq comparing mesenchymal stem cells co-cultured with megakaryocytes

ABSTRACT: RNA-seq comparing mesenchymal stem cells co-cultured with megakaryocytes

PROVIDER: PRJNA1093114 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28491542_1.fastq.gz	Fastqsanger.gz
	SRR28491542_2.fastq.gz	Fastqsanger.gz
	SRR28491543_1.fastq.gz	Fastqsanger.gz
	SRR28491543_2.fastq.gz	Fastqsanger.gz
	SRR28491544_1.fastq.gz	Fastqsanger.gz

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RNA-seq comparing mesenchymal stem cells co-cultured with megakaryocytes

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Project description:Whole livers were collected from mouse fetuses at embryonic day 13.5 (E13.5), and single-cell suspensions were prepared by successive passage through 25-gauge needles. Fetal liver cells were maintained in RPMI1640 (Wako, Osaka, Japan) supplemented with 20% charcoal-stripped fetal bovine serum (FBS), 100 U/ml penicillin, 100µg/ml streptomycin, and 50ng/ml recombinant human thrombopoietin (TPO) (generously provided by Kyowa Hakko Kirin Co. Ltd.). Megakaryocytes were harvested for RNA purification from a day-3 culture. CD41+ cells were enriched using biotinylated anti-CD41 antibody (Serotec; clone MWReg30) and streptavidin-coupled Dynabeads (Dynal Biotech ASA) from a culture of E13.5 fetal liver cells of WT and p45–/– mice. Total RNA was purified from the sorted CD41+ cells. Gene expression was measured in primary megakaryocytes cultured from p45-/- fetal livers and wild type fetal livers at E13.5. Three independent experiments were performed.

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