title: GSM3371744: MF-DC; Mus musculus; RNA-Seq
library_construction_protocol: Mouse hearts were isolated and enzymatically digested in DMEM containing collagenase I (450 U/mL), DNase I (60 U/mL), and hyaluronidase (60 U/mL) and 1mM Flavopiridol in the digestion buffer. Digestions were stopped after 20min and cells were processed into a single cell suspension on ice. Cells were stained with CD45 magnetic beads (Miltenyi Biotec) and subsequently fluorescently tagged antibodies against CD45, CD64 and CD11b. Hematopoietic cells were positively enriched using the AutoMacs instrument (Miltenyi Biotec) and live MFs (CD45+, CD64+, CD11b+, DAPINeg) were sorted on the Aria Fusion (BD Bioscience) under low pressure into DMEM containing 50% FCS  10X single cell RNA sequencing.Cardiac MFs were identified as live, CD45+CD64low-hiCD11b+ and DCs as live,  CD45+CD64-CD11c+MHCIIInt-high.   Libraries were prepared as outlined by the Chromium Single Cell 3' Reagent Kits v2 user guide with appropriate modifications to the PCR cycles based on the calculated cDNA concentration (as recommended by 10X Genomics).
attributes.tag: GEO Accession
attributes.value: GSM3371744