characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-01-31T00:19:46Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-01-31T00:19:46Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110644417
characteristics.characteristic: External Id
characteristics.text: Department of Biosciences, University of Milan and INGM (Istituto Nazionale di Genetica Molecolare), Milan, Italy
characteristics.characteristic: INSDC center name
characteristics.text: 2023-01-31T00:19:46Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-01-31T00:19:46Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12742183
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12080:H9_dif5_d25
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: H9
characteristics.characteristic: cell_line
characteristics.text: striatum neuron
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: At day 15 and day 25 of differentiation, cells were dissociated using Accutase for 5 minutes and filtered through a 20μm cell strainer (Sysmex) to remove cell clumps. Single cell suspensions were counted and adjusted to 1.000-1.200 cells/µl in PBS containing 0.04% Bovine Serum Albumin. 5.000 cells per sample and for each time point were collected for generating Single Cell 3’ Gene Expression libraries. Human ES H9 cell line (WiCell) was cultured on CultrexTM (120-180 μg/mL, Trevigen) -coated dishes in complete mTeSRTM1 medium (STEMCELL TM Technologies) and maintained for a maximum of three months. Medium was changed daily, and cells were dissociated twice a week with PBS (Euroclone) plus 0.5 mM EDTA for passaging or Accutase (Millipore) for 2D striatal differentiation. For neural induction, H9 hESCs cells were plated at a density of 1,1x104 cells/cm2 on CultrexTM- coated plates (120-180 μg/mL) in complete mTeSRTM1 medium supplemented with 10uM ROCK inhibitor (Y-27632, provided by CHDI FOUNDATION). After two days of expansion, cells were exposed to Dual-SMAD inhibition using 10uM SB431542 (provided by CHDI FOUNDATION) and 500nM LDN193189 (provided by CHDI FOUNDATION) in DMEM-F12 (Life Technologies) plus N-2 Supplement (Life Technologies) and B-27 Supplement without Retinoic Acid (Life Technologies) for 12 days. Starting on Day 5, 200 ng/mL Recombinant human SHH C-25 II (R&D SYSTEM) and 100 ng/mL DKK-1 (PrepoTech) were added to the culture and maintained until Day 25. On Day 21 the entire cell population was detached upon Accutase single cell dissociation and replated at a cell density of 2.2x104 cells/cm2 either on Poly-D-Lysin (33 μg/mL, Sigma-Aldrich) and Laminin (3μg/mL, Life Technologies) or Biolaminin 521 (5 μg/mL, Voden) coated plates for continued culture. The cells were maintained, in DMEM-F12 plus N-2 Supplement, B-27 Supplement with retinoic acid (Life Technologies), and 20 ng/mL BDNF (PrepoTech,), until the end of differentiation. GEM generation, reverse transcription, cDNA amplification, and library construction steps were performed according to manufacturer’s instructions using the Chromium Single Cell 3′ v2/v3 platform (10X Genomics) GEM generation, reverse transcription, cDNA amplification, and library construction steps were performed according to manufacturer’s instructions using the Chromium Single Cell 3′ v2/v3 platform (10X Genomics)
characteristics.characteristic: description
characteristics.text: embryo stage
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: striatal differentiation
characteristics.characteristic: growth condition
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: inner cell mass
characteristics.characteristic: organism part
characteristics.text: embryonic stem cell
characteristics.characteristic: progenitor cell type
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: H9_dif5_d25
characteristics.characteristic: title