scRNAseq of human embryo-derived trophoblast organoids and trophoblast stem cells
scRNAseq of human embryo-derived trophoblast organoids and trophoblast stem cells
Trophoblast organoids were cultured at 37 ºC in 21% O2 and 5% CO2 in TOM. Fresh medium was added every two to three days. When organoids reached a size of ~500 µm they were released from Matrigel domes with Dispase for 20 min at 37 ºC and mechanical disruption by pipetting. Organoids were pelleted at 500 x g for 4 min at 4 ºC, resuspended in TrypLE Express and dissociated for 15 min at 37 ºC. Cells were counted and 10,000 cells were seeded per Matrigel dome on a 24-well plate.  For trophoblast stem cells, when cultures reached ~80% confluency, cells were dissociated with TrypLE for 15 min at 37 ºC, pelleted at 500 x g for 4 min  Nucleic extraction was conducted using the Chromium Next GEM Single Cell 5' Reagent Kit v2 (PN-1000263, 10X Genomics) according to the manufacturer's instructions Nucleic extraction was conducted using the Chromium Next GEM Single Cell 5' Reagent Kit v2 (PN-1000263, 10X Genomics) according to the manufacturer's instructions