GSM8003791: Mix of Lin- cells , Non-exposed and HFD-exposed mice, sublibrary 2, multiplexed scRNASeq; Mus musculus; RNA-Seq
Femur bones from HFD-exposed and non-exposed Ldlr-/- mice (n=4/group) were dissected. Bone marrow cells were flushed with DMEM + 10%FCS, washed once and passed through 70 um cell strainers. CD11b+ cells were first depleted with CD11b magnetic microbeads (Miltenyi Biotec, Bergish Gladbach, Germany) and flow through was collected. The cells were later applied to the mouse lineage cell depletion kit (Miltenyi Biotec) to enrich progenitor cells. On average, around 9x105 Lin- cells were isolated per mouse. Lin- cells were fixed according to Evercode Fixation (ParseBiosciences, Seattle, USA) protocol Library preparation was performed following manufacturer's instructions for the Single Cell Whole Transcriptome Mega Kit, where total 12500 cells per sample were projected to be barcoded. After the third round of barcoding, eight sublibraries were prepared with the barcoded cDNA. SPLIT-Seq based Evercode scRNA-Seq