title: TFAP2A KO EB 18h
organism: Homo sapiens
description: Protocols: Embryoid bodies were dissociated using TrypLE, neutralised using MEF media and sorted to remove doublets and debries using a SONY SH800 sorter For the TFAP2A knockout and parental control line E-MTAB-11305 we used HUES9 iCRISPR and H9-TFAP2A lines ESC (https://www.pnas.org/doi/10.1073/pnas.1708341114#sec-3) which are derived from H9 cell line https://www.wicell.org/home/stem-cells/catalog-of-stem-cell-lines/wa09.cmsx. Embryoid bodies were aggregated from PreME cells and treated with BMP to induce PGCLC fate according to Kobayashi et al 2017. PreME/ME protocols were optimised based on previous conditions. For the basal medium, we used aRB27 medium which is composed of Advanced RPMI 1640 Medium (Thermo Fisher Scientific) supplemented with 1% B27 supplement (Thermo Fisher Scientific), 0.1 mM NEAA, 100 U/ml Penicillin- 0.1 mg/ml Streptomycin, 2 mM L-Glutamine. For PreME/ME induction, trypsinized Conv-hPSC were seeded on vitronectin-coated dish at 200,000 cells/well in 12 well plate and cultured in PreME/ME induction medium for 6-36 h as indicated in the figures. PreME/ME induction medium was composed of aRB27 medium supplemental with 100 ng/ml Activin-A (SCI), 3 ŒºM GSK3i and 10 ŒºM of ROCKi. Single Cell 3' 10X V3 Single Cell 3' 10X V3
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-14
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-14
attributes.tag: External Id
attributes.value: SAMEA13409635
attributes.tag: INSDC center alias
attributes.value: The Gurdon Institute, University of Cambridge
attributes.tag: INSDC center name
attributes.value: The Gurdon Institute, University of Cambridge
attributes.tag: INSDC first public
attributes.value: 2023-01-14T00:25:14Z
attributes.tag: INSDC last update
attributes.value: 2023-01-14T00:25:14Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11305:TFAP2A KO EB 18h
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: H9 TFAP2A KO
attributes.tag: cell_type
attributes.value: primordial germ cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: embryo
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: Crspr/Cas9 mediated TFAP2A knockout
attributes.tag: growth condition
attributes.value: embryoid body
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: inner cell mass
attributes.tag: progenitor cell type
attributes.value: embryonic stem cell
attributes.tag: sample name
attributes.value: E-MTAB-11305:TFAP2A KO EB 18h