Homo sapiens
Protocols: The human stem cell clone H9 was grown under previously published conditions. In brief, the hESCs wild-type H9 (WA-09), SATB1KO, NURR1-GFP reporter line were maintained using E8-essential medium (Fisher Scientific) without feeder on VTA-N (Fisher Scientific) and passaged every 4-5 days by EDTA. The knockout of the SATB1 gene was performed by using a directed CRISPR/Cas9 approach in the human stem cell line H9. Stem cell clones were screened by Sanger sequencing to validate SATB1-KO. H9 clones were then differentiated into dopaminergic neurons or cortical neurons (predominantly layer V). Midbrain dopamine (mDA) differentiation from hESC was done with an optimized protocol from previously published by our group (Chambers et al., 2009; Kriks et al., 2011). Cortical differentiation from hESC was done with an optimized protocol from previously published by our group (Maroof et al., 2013). References: Chambers, S.M., Fasano, C.A., Papapetrou, E.P., Tomishima, M., Sadelain, M., and Studer, L. (2009). Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27, 275-280. Kriks, S., Shim, J.W., Piao, J., Ganat, Y.M., Wakeman, D.R., Xie, Z., Carrillo-Reid, L., Auyeung, G., Antonacci, C., Buch, A., et al. (2011). Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson's disease. Nature 480, 547-551. Maroof, A.M., Keros, S., Tyson, J.A., Ying, S.W., Ganat, Y.M., Merkle, F.T., Liu, B., Goulburn, A., Stanley, E.G., Elefanty, A.G., et al. (2013). Directed differentiation and functional maturation of cortical interneurons from human embryonic stem cells. Cell Stem Cell 12, 559-572. RNA was extracted using the RNeasy micro plus kit (Qiagen) according to the producers protocol. The DNA library construction protocol for RNA-Seq was performed using the Illumina TrueSeq RNA sequencing kit.
ENA-FIRST-PUBLIC
2019-07-21T04:02:15Z
ENA-LAST-UPDATE
2018-08-24T19:17:51Z
External Id
SAMEA4858685
INSDC center name
The Rockefeller University Laboratory of Molecular and Cellular Neuroscience
INSDC first public
2019-07-21T04:02:15Z
INSDC last update
2018-08-24T19:17:51Z
INSDC status
public
Submitter Id
E-MTAB-59651535134514:BK19
broker name
ArrayExpress
cell line
H9
cell type
cortical neuron
common name
human
genotype
NURR1::GFP; SATB1 knockout
progenitor cell type
embryonic stem cell
sample name
E-MTAB-59651535134514:BK19
scientific_name
Homo sapiens