biostudies-arrayexpressHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-13862Transcriptome of young HSCs (Lin-CD34+CD38-), young Progenitors (Lin-CD34+CD38+) and old HSCs (Lin-CD34+CD38-) isolated from bone marrow samples of healthy donors.biostudies-arrayexpressLibrary Construction - cDNA libraries were generated using SMARTseq v4 (Takara Bio). For HSCs, 13 cycles of amplification were performed. To produce uniquely- and dually-barcoded sequencing libraries from the cDNA libraries, the NEBNext Ultra II FS DNA library kit (NEB) was utilized. This involved fragmenting 5 ng of the cDNA library for 22.5 minutes, followed by adaptor ligation and library amplification using cycle numbers determined by amount of input material.Nucleic Acid Extraction - Cells were sorted into RNA lysis buffer (Arcturus PicoPure RNA Isolation Kit (Applied Biosystems)) and then stored at -80οC until further use. RNA isolation was conducted using the Arcturus PicoPure RNA Isolation Kit (Applied Biosystems) following the manufacturer's guidelines. DNAse treatment was carried out employing the RNase-Free DNase Set (Qiagen). The resulting total RNA was utilized for generating cDNA libraries.Sequencing - Libraries underwent sequencing on the Illumina NovaSeq platform, generating 50 million reads depth with 100bp paired-end sequencingSample Collection - Human frozen bone marrow samples were thawed at 37oC and transferred to 10ml of thawing media (IMDM, 10% FCS, 1mM EDTA, 1:1000 DNase) and spun down for 8 min at 300g. Cells were resuspended in Stem Span and let for recovery in the incubator for 20 min. Cells were resuspended in 500ul antibody mix (Lin(CD3/14/16/19/20/56)-APC; CD38-Pe/Cy7, CD34-FITC*)  at 4oC for 45 min. Cells were washed with 0.9% NaCl and incubated with Zombie-Aqua (1:1000) for 10 min at RT. Cells were finally washed and resuspended in 0.9% NaCl and filtered for FACS sorting. Sorting was then performed using a FACS Aria III at the MPI FACS facility, Freiburg, Germany. 1000 cells of young HSCs/Progenitors and old HSCs were sorted in 4 and 7 biological replicates, respectively.MINSEQE ScoreAssays and DataorganisationMAGE-TAB FilesMetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNAHomo sapiensE-MTAB-13863Maria-Eleni LaliotiMari Carmen Romero-MuleroNoemie KarabaczNina Cabezas-WallscheidKarin JaeckleinfalsePopulation RNA-seq of young hematopoietic stem cells (HSCs) vs young progenitors vs old HSCs isolated from human bone marrowTranscriptome of young HSCs (Lin-CD34+CD38-), young Progenitors (Lin-CD34+CD38+) and old HSCs (Lin-CD34+CD38-) isolated from bone marrow samples of healthy donors.2025-05-20T00:00:00Z2026-06-05T16:50:40.406Z2024-02-27T22:43:56.579ZE-MTAB-13862ERP158168E-MTAB-13863EFO_0002944EFO_0004170EFO_0005518EFO_0003738EFO_0004184