2024-09-18
ENA first public
ERC000011
ENA-CHECKLIST
BAH-GU
INSDC center name
public
INSDC status
ERS21049989
SRA accession
E-MTAB-14450:12886_Holstein
Submitter Id
5
year
age
ArrayExpress
broker name
macrophages
cell_type
not collected
collection date
domestic cattle
common name
Protocols: Two Theileria infected cell lines were utilised. One line (Sah1 (82H)) represented an ex vivo isolate derived from an infected calve of the Sahiwal breed (Bos indicus) while the other (Hol 3 (12886)) was derived from an infected Holstein calf (Bos taurus).Both Sahiwal and Holstein animals were infected with sporozoites from the same, T. annulata Hissar stock and cultured as standard in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% FCS, 4 mM L-glutamine and 50 μM β-mercaptoethanol, at 37 0C for a limited period (l2 passages). Both cell lines were fully established based on the level of infection (>95% macroschizont infected cells). Cultures were set up at 1X105cells/ml and cultured for 48 hours. Cell counts and viability were determined by trypan blue exclusion using a CountessTM 3FL automated cell counter (Invitrogen). Cell viability was 94% (Sah 1) and 95% (Hol 3), while growth for the Holstein line was 1.58 fold greater than the Sahiwhal. The two different bovine species were infected with Theileria annulata Multiome nuclei extraction protocol was adapted from(dx.doi.org/10.17504/protocols.io.rm7vzyx75lx1/v1), cells were harvested by centrifugation (400 g for 10 min), and lysed for 10min on the rotor at 4 ̊C with 1X CST lysis buffer (as indicated in dx.doi.org/10.17504/protocols.io.rm7vzyx75lx1/v1). Lysate was filtered with 40 μm mini cell strainer into a clean pre-labelled 5.0 mL Eppendorf tube and PBS + 1% BSA was added to dilute the lysis buffer. The tube was spun down 500g, 4°C, 5 min. Nuclei were resuspended and counted with Neubauer haemocytometer; 10,000 nuclei were loaded for each chip line on a 10X Genomics Multiome J chip (#1000230).  Libraries were prepared according to the 10X Genomics multiome protocol (#1000285, #1000215, #1000212)
description
ex vivo
developmental stage
theileriasis
disease
unknown
genotype
not collected
geographic location (country and/or sea)
12886
individual
not applicable
isolate
Bos taurus
organism
Bos taurus
organism
macrophages
organism part
susceptible
phenotype
Bos taurus
scientific_name
male
sex
Holstein
strain
12886_Holstein
title