Single-cell RNA-sequencing data from developing embryonic human finger joints
Single-cell RNA-sequencing data from developing embryonic human finger joints
Skin and tendon material were removed from each finger, and DIP and PIP joints isolated by cutting either side of the joint space. Joints were cut into pieces and enzymatically digested with 1mg/ml collagenase D in DMEM, at 37 degrees C for 1h 30mins. Supernatant containing released cells was collected every 10-15 mins, neutralized in DMEM 10% FBS, and stored at 4 degrees C. Once supernatant was removed, fresh enzyme was applied to continue sample digestion. After digestion, the remaining tissue was manually dissociated, and combined with collected supernatants, in DMEM 10% FBS. All isolated cells were then passed through a 70μm filter. Where possible, cells were sorted for viability using the 7-AAD dye (Biolegend 420403) and loaded onto the 10X Genomics Chromium Controller (Chip G) following the manufacturer's protocol to capture 10,000 cells per reaction.  As per 10x Genomics Single Cell 3' Reagent Kits v3.1 manufacturer user guide (CG000330 Rev B) As per 10x Genomics Single Cell 3' Reagent Kits v3.1 manufacturer user guide (CG000330 Rev B)