title: Illumina HiSeq 2500 paired end sequencing; Single-cell RNA-seq of of dermal fibroblasts using microfluidic droplet capture (unstimulated cells)
design: Single-cell RNA-seq of of dermal fibroblasts using microfluidic droplet capture (unstimulated cells)
library_name: Sample 555_p
library_construction_protocol: Cells were washed with PBS and then trypsinized.  Human dermal fibroblasts were obtained from the HipSci consortium (http://www.hipsci.org/ (Streeter et al., 2017) (where they were used for creating iPSCs). These original cells were thawed to test for growth and viability and refrozen after 1-2 passages, so that all lines were in the same growth rate and proliferation conditions before the stimulation experiment. A mix of 3 cell lines (HPSI1013pf-wuye, HPSI1113pf-wetu, HPSI0114pf-vass) was used. Cells were grown in DMEM, high glucose, GlutaMAX Supplement, pyruvate (Life Technologies / 10569-010) + Fetal Bovine Serum (Lab Tech / FB-1001) + Penicillin – Streptomycin (Life Technologies / 15140122).  unstimulated control Cells were directly processed in a 10X Chromium machine (10X Genomics) according to the manufacturer's protocol. The Single Cell Gel Bead kit (#120217), Single cell chip kit (#120219) and Single cell library kit (#120218) were used along with 10x GemCode Single Cell Instrument as per manufacturer specifications and manuals (Document # CG00011; Revision B).
attributes.tag: Experimental Factor: single cell identifier
attributes.value: cell 555