Homo sapiens
Protocols: For harvesting and generating single cell suspensions, human iPSCs were harvested using Accutase (Millipore, product code SCR005). Just prior to FACS sorting, cells were resuspended in 1x PBS and strained (Cell strainer (40uM), Corning #CLS431750-50EA). Single cells were sorted into 96-well plates (Framestar #4TI-0960) containing 2 µl lysis buffer with 1U/µl Rnase Inhibitor (RRI), ERCC spike in mix at 1 in 20 Mio dilution (Ambion Cat #4456740). Plates were sealed (Thermo Fisher Scientific #AB0626) immediately spun down and frozen at −80 °C until further processing. no growth protocol Human-induced pluripotent stem cells were thawed and cultured under feeder-free conditions on vibronectin coated 15 cm tissue culture treated plates (Corning) in complete medium (500ml Essential 8 medium ((Life Technologies #A1517001), 10 ml Essential 8 supplement, 5ml Penicillin/Streptomycin (Invitrogen #15140122)). Cells were media changed every day except for the day of passaging. Cells were incubated at 37°C, 5% CO2 and routinely passaged using 0.5 mM EDTA (LifeTechnologies #AM9260G). no treatment We prepared cDNA of human iPSCs following the Smart-seq2 protocol.
ENA-FIRST-PUBLIC
2020-10-01T17:06:44Z
ENA-LAST-UPDATE
2020-09-29T17:06:25Z
External Id
SAMEA7364985
INSDC center name
Cambridge University CRUK-CI
INSDC first public
2020-10-01T17:06:44Z
INSDC last update
2020-09-29T17:06:25Z
INSDC status
public
Submitter Id
E-MTAB-9577:hipsci_shortread_85
broker name
ArrayExpress
cell type
induced pluripotent stem cell
common name
human
genotype
wild type genotype
individual
HPSI-euts
sample name
E-MTAB-9577:hipsci_shortread_85
scientific_name
Homo sapiens
sex
male