characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-02-23T00:28:36Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-02-23T00:28:36Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112642719
characteristics.characteristic: External Id
characteristics.text: Department of Hematology Erasmus MC
characteristics.characteristic: INSDC center name
characteristics.text: 2023-02-23T00:28:36Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-02-23T00:28:36Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14634688
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12616:BL6_d21_B_T_Mono_Neutro_3
characteristics.characteristic: Submitter Id
characteristics.text: 12
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: bone marrow cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: 1 femur and 2 tibias from each mouse were used. After isolating and cleaning the bones, one end was cut off and bones were placed into a 0.5 ml microcentrifuge tube with a hole punched in the bottom. This was nested into a 1.5 ml microcentrifuge tube and spun down at 10,000 RPM for 15 seconds to collect the bone marrow. Following resuspension in PBS containing 1% FCS and 1 mM EDTA, erythrocytes were lysed. Remaining cells were washed with PBS containing 2% FCS, filtered through a 100 um cell strainer and counted. Following erythrocyte lysis, the samples were filtered through a 100 um cell strainer and counted. Fc-receptors were blocked with 10% normal mouse serum and anti-CD16/32 (BD, clone 2.4G2) prior to incubation with the following antibodies: CD3e-PerCpCy5.5 (eBioscience, clone 145-2C11), CD8-FITC (BD, clone 53-6.7), CD11b-APCeFluor780 (eBioscience, clone M1/70), CD19-AF700 (eBioscience, clone eBio1D3), CD27-BV510 (Biolegend, clone LG.3A10), CD44-BV510 (Biolegend, clone IM7), NKp46-PE-Cy7 (eBioscience, clone 29A1.4), Gr1-APC (BD, clone RB6-8C5), Ly6c-BV510 (Biolegend, clone HK1.4). Samples were incubated for 25 minutes on ice in the dark. Following staining, cells were washed and resuspended in PBS + 2% FCS containing DAPI, and sorted with a BD FACS Aria III. Sorted T, NK, B, monocyte/macrophages and neutrophil granulocytes were combined per mouse and processed as one sample. Single cells were encapsulated for cDNA synthesis and barcoded using the Chromium Single Cell 3' Reagent Kit v3 (10X Genomics) Libraries were constructed according to the manufacturer’s recommendations (10X Genomics)
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: multiple myeloma
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: B_mm_3
characteristics.characteristic: individual
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: bone marrow
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: C57Bl/6JOlaHsd
characteristics.characteristic: strain
characteristics.text: BL6_d21_B_T_Mono_Neutro_3
characteristics.characteristic: title