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SRX23732156: GSM8107936: Second_kidney_xeno_scRNA_seq_untransplanted; Sus scrofa; RNA-Seq
8 ILLUMINA (Illumina NovaSeq 6000) runs: 380.4M spots, 48.3G bases, 14.9Gb downloads

External Id: GSM8107936_r1
Submitted by: Jef Boeke, Institute for Systems Genetics, NYU Langone Health
Study: Cellular dynamics of pig-to-human kdiney xenotransplantation [single cell RNA-seq]
show Abstracthide Abstract
We performed comprehensive single-cell RNA sequencing (scRNA-seq) analyses of the two porcine xenografts and their contralateral untransplanted kidneys to dissect xenotransplantation-associated cellular dynamics and xenograft-recipient interactions. We additionally performed longitudinal bulk RNA-seq on core biopsies to dissect the time-resolved physiological changes of the xenografts and longitudinal scRNA-seq of peripheral blood mononuclear cells (PBMCs) to detect recipient's immune responses across time. Overall design: Xenografts and contralateral control kidneys went through the same procedure for preparation of transplantation. Kidney tissues from xenograft and control kidneys in both cases were collected at the end of the study (54 hours). PBMC samples were collected from the recipient in case 2 at the following timepoints: pre-transplant (0 h) and then at 6h, 12h, 24h, 48h, and 53h (right before termination).
Sample: Second_kidney_xeno_scRNA_seq_untransplanted
SAMN40120828 • SRS20560380 • All experiments • All runs
Organism: Sus scrofa
Library:
Name: GSM8107936
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC SINGLE CELL
Selection: cDNA
Layout: PAIRED
Construction protocol: Fresh kidney tissues were minced rapidly in a cell culture dish followed by washing with DPBS buffer. Minced tissue was resuspended in 5mL of 37°C pre-warmed 0.25% Trypsin for 20 min with mechanical dissociation using serological pipette every 5 min. Following digestion, dissociation mix was filtered through a 100µm strainer. The remaining large tissues on the strainer were further ground and washed to collect additional cells. Cell debris and dead cells were removed using Miltenyi Biotech Dead Cell Removal Kit before loading for 10X Genomics single-cell RNA-seq (v3.1 Chemistry). Longitudinal single-cell RNA-seq of the recipient's PBMCs were prepared at the same time after collecting all samples across timepoints. Fresh PBMCs of each timepoint were first collected through gradient centrifugation, followed by cryopreservation of the prepared PBMCs according to the 10X Genomics protocol (protocol number: CG00039). Upon collecting all PBMC samples, the frozen cells were thawed at the same time, then loaded for 10X Genomics single-cell RNA-seq (v3.1 Chemistry). prepared signle cells were sequenced using 10X Genomics single-cell RNA-seq (v3.1 Chemistry) following manufacturer's protocol.
Runs: 8 runs, 380.4M spots, 48.3G bases, 14.9Gb
Run# of Spots# of BasesSizePublished
SRR2808566854,565,6336.9G2.1Gb2024-06-30
SRR2808566955,131,4547G2.1Gb2024-06-30
SRR2808567051,578,3996.6G2Gb2024-06-30
SRR2808567152,139,8866.6G2Gb2024-06-30
SRR2808567241,274,8405.2G1.6Gb2024-06-30
SRR2808567341,611,6595.3G1.6Gb2024-06-30
SRR2808568841,775,8595.3G1.6Gb2024-06-30
SRR2808568942,302,4585.4G1.7Gb2024-06-30

ID:
32021845

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