|
| Status |
Public on Feb 27, 2024 |
| Title |
iPSC derived sensory neurons |
| Sample type |
SRA |
| |
|
| Source name |
BJ-iPSC
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: BJ-iPSC
cell type: Induced pluripotent stem cell-derived Sensory neurons
genotype: wildtype
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
For cell detachment, 46-day old hiPSC-SNs were treated with accutase for 8 min at 37°C and transferred into 15 mL falcon tube before topping up with dPBS to 10 mL and centrifuging at 1000 rpm for 3 min. Supernatant was discarded and hiPSC-SNs were dissociated using papain (Worthington Biochemical) according to manufacturer’s instructions. Briefly, papain digestion was carried out at 37°C with constant agitation for 30 min and with manual pipetting done every 10 min. Cells were spun down at 1000 rpm for 3 min and the supernatant was discarded. The reaction was stopped by re-suspension and incubation in diluted DNase/albumin-inhibitor solution at 37°C for 5 min before spinning down the cells again at 1000 rpm for 3 min and discarding the supernatant. Finally, dissociated cells were resuspended into 500 μL Sensory neuron maturation medium to a concentration of ~ 320 cells/μL and filtered through a 40 µm cell strainer to remove any remaining undissociated cell clumps.
The single-cell suspensions were assessed by acridine orange/propidum iodide staining to contain a cell viability of 97.2%. Using the GEXSCOPE kit, the cells were loaded into microwells of a microfluidic SCOPE-chip and lysed to release mRNA hybridizing to unique barcode sequences on the bead in the same well. The hybridized mRNA was reverse transcribed into cDNA and amplified. The resulting amplified cDNA was fragmented, ligated to adaptors and processed to construct a sequencing library compatible with the Illumina sequencing platform.
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic single cell |
| Library selection |
cDNA |
| Instrument model |
Illumina NovaSeq 6000 |
| |
|
| Description |
Libraries were 150PE sequenced on the Illumina Novaseq platform
|
| Data processing |
Raw sequencing reads were processed by Celescope pipeline RNA assay with default parameters to generate filtered single-cell gene expression matrix files (https://github.com/singleron-RD/CeleScope).
Assembly: Homo_sapiens_ensembl_99
Supplementary files format and content: Expression Matrix
Supplementary files format and content: Cell Barcodes
Supplementary files format and content: Gene Information
|
| |
|
| Submission date |
Feb 23, 2024 |
| Last update date |
Feb 27, 2024 |
| Contact name |
Shi-Yan Ng |
| E-mail(s) |
syng@imcb.a-star.edu.sg
|
| Organization name |
A*STAR
|
| Street address |
61 Biopolis Drive
|
| City |
Singapore |
| ZIP/Postal code |
138673 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL24676 |
| Series (1) |
| GSE256435 |
iPSC-derived human sensory neurons reveal a subset of TRPV1 antagonists as anti-pruritic compounds |
|
| Relations |
| BioSample |
SAMN40007502 |
| SRA |
SRX23677916 |
