characteristics.text: 2025-01-08
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: KU Leuven
characteristics.characteristic: INSDC center name
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS21186774
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-14067_3:EXT352
characteristics.characteristic: Submitter Id
characteristics.text: 24 to 26
characteristics.unit: month
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: Anterior pituitary cells
characteristics.characteristic: cell_type
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Mice were euthanized by CO2 asphyxiation, followed by decapitation. Heads were washed and sprayed with 70% ethanol. Head skin was removed, the cranium opened and the brain withdrawn. For the -/iDTR and GhCre/iDTR mice, the neurointermediate lobe was discarded under a stereomicroscope. The (anterior) pituitary (AP) was collected in medium A. Supernatant was removed and the AP incubated with prewarmed trypsin (2.5%; Gibco) for 15 minutes at 37°C. Then, DNase (2 µg/mL in medium A; Sigma) was added and the suspension gently mixed. Supernatant was removed and the suspension incubated with trypsin inhibitor (1 mg/mL in medium A; Sigma) for 10 minutes at 37°C. Supernatant was removed, medium B1 was added, and samples were incubated at 37°C. After 5 minutes, medium C was added, and the suspension was again incubated at 37°C. Supernatant was removed after 15 minutes and fragments were washed with medium C. The AP fragments were then mechanically dissociated using a flame-polished Pasteur pipet and transferred to medium A with DNase (2 µg/mL). The resulting suspension was filtered through a 40 µm cell strainer; 3% BSA (Serva) was layered at the bottom of the recipient and the sample centrifuged for 10 minutes at 190 x g. Supernatant was removed, the pellet resuspended in 1 mL of Advanced DMEM/F-12, and cells were counted using a NucleoCounter (Chemometec). The cell suspension was centrifuged again (10 minutes at 190g) and resuspended in PBS with 0.04% BSA at a concentration of 1x10^6 cells/mL.  GHCre/iDTR mice and Cre-negative control littermates were intraperitoneally injected with 4 ng DT/g bodyweight (Sigma-Aldrich, Bornem, Belgium) twice a day (8 h in between the injections) for 3 consecutive days. Pituitaries were isolated and analyzed the following day (day 4). The AP single-cell suspensions were converted to barcoded Drop-seq libraries by using the Chromium Single Cell 3’ V3.1 Library, Gel Bead & Multiplex Kit and Chip Kit (10X Genomics; Pleasanton, California, USA), aiming for an estimated 8,000 cells per library. Single-cell libraries were prepared with the Chromium Single Cell 3’ V3.1 Chemistry Library Kit, Gel Bead & Multiplex Kit and Chip Kit from 10X Genomics.
characteristics.characteristic: description
characteristics.text: mouse postnatal
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: GhCre/iDTR
characteristics.characteristic: genotype
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: EXT352
characteristics.characteristic: individual
characteristics.text: endocrine cell-ablation injury
characteristics.characteristic: injury
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Pituitary gland
characteristics.characteristic: organism part
characteristics.text: Anterior pituitary
characteristics.characteristic: sampling site
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: EXT352
characteristics.characteristic: title