GSM8794755: HSC-LO_Day10; Homo sapiens; RNA-Seq
The differentiated hiPSC-HSC_P0  were detached from culture well for scRNA sequencing analysis. For HSC-LO_Day10 dissociation: After 10 days of co-culturing hiPSC-hepatic endoderm, hiPSC-endothelial cells, and hiPSC-HSCs, the liver organoids(LOs) were washed with PBS and collected in one well. 2mL of the dissociation medium (HBSS, 1%HEPS, 0.5%CaCl2, 0.5%MgCl2, and 3mg/mLCOIIV) was added into one well of 6-well plate, incubate the LOs at 37℃ for 30min. Asparite the dissociation medium and add 2mL/well(6-well plate) TrpLE and incubate LOs at the room temperature for 10min. Dissociate LOs by using pippete and collect cells in washing medium (DMEM supplemented with 1% PS and 10% FBS)  The collected cells were filtered through a 100 μm cell strainer and centrifuged at 200 g for 4 minutes. Subsequently, CD31-APC antibody (Miltenyi Biotec) and anti-APC MicroBeads (Miltenyi Biotec) were employed to enrich the endothelial cell population for downstream analyses Library was prepared by reagents from a Chromium Single Cell 3'v3 reagent kit, according to the 10x Genomics manual.