2025-10-30
ENA first public
ERC000011
ENA-CHECKLIST
Dipartimento di Biotecnologie cellulari e scienze della salute
INSDC center name
public
INSDC status
ERS25142705
SRA accession
E-MTAB-15308:barcode_I
Submitter Id
ArrayExpress
broker name
WTC11
cell line
hPSC derived cortical neuron organoids
cell type
not collected
collection date
human
common name
Protocols: Human neural organoids were cultured for 30 days following the protocol outlined by Velasco et al. (2019) PMID: 31168097. iPS2seq transfection pools were differentiated separately, and tetracycline was added to the treated organoids throughout the culture. Organoids were dissociated into single-cell suspensions using papain-based enzymatic digestion as described in Protocol Exchange (https://protocolexchange.researchsquare.com/article/pex-258/v1). Cell counts and live cell estimation were determined using a hemocytometer, and suspensions were adjusted to 2 million cells per tube. Then cells were labeled with CMO, sorted for vital cells and then loaded on the 10X controller. Cell Multiplexing Oligo (CMO) labeling was performed according to the manufacturer’s protocol (CG000391) with an additional washing step. Cells were pooled in equal quantities, and the cell-pooled suspension was counted to ensure a final concentration of 1,600 cells/μL to obtain a final cell recovery of 30000 cells. Single-cell libraries were prepared using the Chromium Next GEM Single Cell 3' v3.1 protocol (CG000388 Rev C). Cells were loaded on a chip G. CG000388 Rev B (with cell multiplexing oligos). hPS-2seq barcode enrichment protocol was performed with custom primers binding on the TET repressor.
description
DD35
developmental stage
normal
disease
Barcodes KD screening in pool
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
cortical neuron organoid
organism part
Homo sapiens
scientific_name
barcode_I
title