ERC000011
ENA-CHECKLIST
2023-11-30T00:25:00Z
ENA-FIRST-PUBLIC
2023-11-30T00:25:00Z
ENA-LAST-UPDATE
SAMEA113447829
External Id
Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases Department of Biosciences University of Milan and Istituto Nazionale di Genetica Molecolare
INSDC center name
2023-11-30T00:25:00Z
INSDC first public
2023-11-30T00:25:00Z
INSDC last update
public
INSDC status
ERS15442932
SRA accession
E-MTAB-12924:Sample 13
Submitter Id
ArrayExpress
broker name
RUES2
cell_line
neuronal stem cell
cell_type
human
common name
Protocols: Additional protocol information can be found in the Addtional Files. The organoids were transferred in a dish to cut away the excess of Matrigel with a blade and then were processed by dissociation using Papain Dissociation System with DNase (Worthington, LK003150) following the manufacturer’s recommendations. The incubation time was adjusted, based on size, at 20 min in the orbital shaker at 37°C. Then the samples were centrifugate for 1min at 1000 rpm and resuspended in DMEM+Neurobasal containing N2 supplement (1:100, Gibco), B27 supplement (1:50, Gibco), and Rock Inhibitor Y-27632 (10µM, provided by CHDI Foundation, CHDI-00197406-0001-007). For the dissociation into single cell suspension, the samples were pipetted up and down first with 1000µl pipettes and following with glass pipettes of increasingly smaller tip diameter (fire-polished). The suspension was filtered with disposable filter of 20µm (Celltrics sysmex, 04-004-2325). The cells were counted with Countess Automated Cell Counter to check viability (higher than 90%) and concentration. The suspension was centrifugated for 1 min at 1000 rpm and resuspended at the concentration of 1x104 cells /µl in PBS containing 0.04% weight/volume BSA (Ambion, AM2616, 400 µg/ml). The Chromium Single Cell 3′ Library & Gel Bead Kit v2 (PN- 120237), Chromium Single Cell 3′ Chip kit v2 (PN-120236) and Chromium i7 Multiplex Kit (PN-120262) were used according to the manufacturer’s instructions in the Chromium Single Cell 3′ Reagents Kits V2 User Guide The Chromium Single Cell 3′ Library & Gel Bead Kit v2 (PN- 120237), Chromium Single Cell 3′ Chip kit v2 (PN-120236) and Chromium i7 Multiplex Kit (PN-120262) were used according to the manufacturer’s instructions in the Chromium Single Cell 3′ Reagents Kits V2 User Guide
description
Huntington disease
disease
CRISPR/Cas9 modification of HD
genotype
brain organoid single culture
growth condition
Homo sapiens
organism
Homo sapiens
organism
inner cell mass
organism part
embryonic stem cell
progenitor cell type
Homo sapiens
scientific_name
Sample 13
title