organism: Homo sapiens
description: Protocols: AML samples were collected from diagnostic peripheral blood (PB) aspirations at the University hospital in Heidelberg in accordance with the Declaration of Helsinki after obtaining written consent from each patient. The project was approved by the Ethics Committee of the Medical Faculty of Heidelberg (S-169/217). PB mononuclear cells (MNCs) were isolated by density gradient centrifugation using Ficoll Paque Plus (GE Healthcare, cat # GE17-1440-03), and stored in liquid nitrogen until further use. Primary AML cells were stained with a total of ten fluorescent cell surface antibodies (see supplementary table X). Cells were sorted into four populations according to CD11b, CD64 and GPR56 expression within the lineage-negative gate. Cells from each population and from lineage-neagtive bulk were sorted directly into RNA extraction buffer (Thermo Fisher, cat # KIT0214), snap-frozen and stored at -80 °C until RNA extraction. RNA extraction and purification of FACS-sorted cells was done using PicoPure RNA Isolation Kit according to manufacturer's instructions (Thermo Fisher, cat # KIT0214). Whole transcriptome amplification was performed using a modified smart-seq2 protocol (Picelli et al 2014), with 5μl of a modified RT buffer containing 1× SMART First Strand Buffer (Takara Bio Clontech, cat # 639538), 1 mM dithiothreitol (Takara Bio Clontech), 1 μM template switching oligo (IDT), 10 U μl−1 SMARTScribe (Takara Bio Clontech, cat # 639538) and 1 U μl−1 RNasin Plus RNase Inhibitor (Promega, cat # N2615). Tagmentation of cDNA was done using Nextera XT DNA Library Preparation Kit (Illumina, cat # FC-121-1030).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-01
attributes.tag: External Id
attributes.value: SAMEA110398840
attributes.tag: INSDC center alias
attributes.value: Division of Stem Cells and Cancer, German Cancer Research Center (DKFZ) and DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM gGmbH), 69120 Heidelberg, Germany
attributes.tag: INSDC center name
attributes.value: Division of Stem Cells and Cancer, German Cancer Research Center (DKFZ) and DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM gGmbH), 69120 Heidelberg, Germany
attributes.tag: INSDC first public
attributes.value: 2023-01-01T00:28:18Z
attributes.tag: INSDC last update
attributes.value: 2023-01-01T00:28:18Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11976:Sample 23
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: acute myeloid leukemia
attributes.tag: disease staging
attributes.value: monocyte-like acute myeloid leukemia
attributes.tag: immunophenotype
attributes.value: CD64+CD11b+GPR56+
attributes.tag: individual
attributes.value: HIAML27
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: blood
attributes.tag: sample name
attributes.value: E-MTAB-11976:Sample 23
attributes.tag: sampling site
attributes.value: leukemic stem cell