GSM4968842: liver 5.5 week; Homo sapiens; RNA-Seq
For single cell RNA-sequencing, dissociated cells were stained with the following antibodies: human-CD45-APC-H7 cl.2D1 (BioLegend 368516; 1:100), CD34-BV605 cl. 581 , CD34-BV605 cl. 581 , CD90-APC cl. 5E10 (BD 562556; 1:100), CD38-PE-Cy7 cl. HIT2 (BD 560677; 1:100), CD43-FITC cl.1G10 (BD 555475; 1:20), CD235a-PE cl. 3H9. CD34+ and/or CD31+ cells were sorted into PBS 0.04% BSA using a BD FACS Aria cell sorter. Placenta, second trimester liver and cord blood were magnetically enriched for CD34+ cells using human CD34 MicroBead Kit UltraPure (Miltenyi) and following manufacturer instructions. For the generation of single-cell gel beads in emulsion, cells were loaded on a Chromium single cell instrument (10x Genomics) with an estimated targeted cell recovery of ~5,000 cells as per manufacturer's protocol. In brief, single-cell suspension of cells in 0.4% BSA-PBS were added to each channel on the 10X chip. Cells were partitioned with Gel Beads into emulsion in the Chromium instrument where cell lysis and barcoded reverse transcription of RNA occurred following amplification. Single-cell RNAseq libraries were prepared by using the Chromium single cell 3' library and gel bead kit v2 (10X Genomics).