TCR_Bl67
Homo sapiens
Protocols: We conducted a single-centre cross-sectional study in adult patients (>18 years of age) undergoing liver transplantation (LTX) for PSC (n=19), ALD (n=16) or resection for PSC-associated cholangiocarcinoma (to which we refer as PSC early-stage due to absence of cirrhosis n=2) at the University Medical Center Hamburg-Eppendorf (UKE, Hamburg, Germany). Liver tissue and whole blood were obtained from each patient at the time of surgery. Diseases were diagnosed and treated following the guidelines of the European Association for the Study of the Liver (EASL). Clinical parameters were recorded immediately before surgery. PSC patients with earlier disease stages (PSC early, n=8) were recruited from our outpatient clinic for phenotyping and functional experiments from peripheral blood. Sex and age matched healthy blood donors (HD, n=10) were used as additional controls. All patients and donors participating in this study provided written informed consent according to the ethical guidelines of the Institutional Review Board of the medical faculty of the University of Hamburg (PV4081 & PV4898). The scTCR-Seq library was prepared using the 10x Single Cell V(D)J Kit v1, according to the manufacturer's instructions. FACS-sorted cells were washed once with PBS containing 0.04% bovine serum albumin (BSA) and resuspended in PBS. GEM generation was performed using the 10x Chromium Controller, cells were loaded on the 10x A Chip according to GEM generation user guide (10x Genomics, USA). Droplet preparation was followed by reverse transcription and cell barcoding, emulsions were resolved and the cDNA was purified using Dynabeads MyOne SILANE (ThermoFisher Scientific, USA) followed by a PCR amplification. Amplified cDNA was then used for enrichment of full-length V(D)J segments (10x Barcoded) via PCR amplification with primers specific to the TCR regions. For TCR library construction, amplified cDNA was fragmented and end-repaired, double-sided size-selected using SPRIselect beads.
ENA first public
2022-04-01
ENA last update
2022-04-01
External Id
SAMEA13856692
INSDC center alias
University Medical Center Hamburg-Eppendorf
INSDC center name
University Medical Center Hamburg-Eppendorf
INSDC first public
2022-04-01T12:11:00Z
INSDC last update
2022-04-01T12:11:00Z
INSDC status
public
Submitter Id
E-MTAB-10143:TCR_Bl67
age
62
batch
10
broker name
ArrayExpress
cell type
naive T cell
common name
human
developmental stage
adult
disease
primary sclerosing cholangitis
individual
20
organism part
blood
sample name
E-MTAB-10143:TCR_Bl67
sex
male