GSM4860673: fresh_HSC40; Mus musculus; RNA-Seq
Haematopoietic Stem Cells (HSCs) were extracted from primary mouse bone marrow, derived from C57BL/6 mice(12-16 weks). For this purpose, lineage depleted bone marrow cells were FACS sorted. HSCs were defined as EPCR-high, CD45+, Sca-1-high, CD48-low/neg, CD150+ (ESLAM). HSCs cultured in hibernated conditions (hibHSCs), hibHSCs stimulted with SCF in culture (hibHSC_SCF), freshly isoalted HSCs (fresh_HSC) and fresh_HSCs stimulated with SCF (fresh_HSC_SCF) were sequenced. Cultured HSCs were FACS sorted to be deposited in 96 well plates (one cell per well), containing lysis buffer and subsequently processed using the previously described Smart-Seq2 protocol (Picelli et al., 2014). Smart-Seq2 protocol performed as previously described (Picelli et al., 2014). Libraries were prepared using Illumina Nextera libraries, single end reads and 50bp read lengths. The pooled libraries were sequenced on an Illumina HiSeq 4000.