GSM4610617: Bone marrow stroma Dll4(E12); Mus musculus; RNA-Seq
Femurs and tibiae were harvested 3 weeks post-injection from adult mice at the age of 11 weeks. Bones were crushed by pestle and mortar with ice-cold 2% FCS/PBS and followed by 30 min digestion at 37 °C with collagenase cocktail by mixing with 2 mg/ml of collagenase IV (17104-019, Gibco), 2 mg/ml collagenase I (17100-017, Gibco), and 10 U/ml of Dispase (17105-041) in a 1:1:1 ratio. The resulting single cell suspension was subjected to depletion of lineage-positive hematopoietic cells with following reagents from Miltenyi Biotec: Lineage Cell Depletion Kit (130-090-858), CD45 Microbeads (130-052-301), CD117 Microbeads (130-091-224), and Ter-119 Microbeads (130-049-901). Lineage-depleted stromal cells were loaded into a BD Rhapsody™ Cartridge for capturing single cells for whole transcriptome according to the manufacturer's instructions. scRNA libraries were prepared with The BD Rhapsody Whole Transcriptome Analysis (WTA) Amplification kit.
GEO Accession
GSM4610617