10x single cell 5MO head
Planarian heads were surgically removed along the posterior end of the auricle.  Tissues were macerated with a surgical knife in CMF buffer supplemented with 0.5% BSA (CMFB) as previously described42. Macerated tissues were kept on ice-cold CMFB in a 6-cm petri dish with gentle horizontal shaking for 20 minutes. Tissues were pipetted up and down every 5 minutes with a transfer pipette 5 to 7 times to ensure sufficient disassociation and optimal viability. Cells were filtered through a 40m strainer before collection by spinning at 400rcf for 5 minutes at 4C. Cells were counted manually. Cell viability was assessed by trypan blue (0.4%). A viability reading of more than 70% is required before library preparation. The single cells were captured on the Chromium Controller (10x Genomics) and libraries were prepared using the Chromium Single Cell 3 Gel Bead and Library Kit (10x Genomics).