characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-03-31T00:22:31Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-03-31T00:22:31Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112484754
characteristics.characteristic: External Id
characteristics.text: Wellcome Sanger Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2023-03-31T00:22:31Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-03-31T00:22:31Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14602101
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12421:Pla_HDBR8624430
characteristics.characteristic: Submitter Id
characteristics.text: 12
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: surgical terminal of pregnancy
characteristics.characteristic: clinical information
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Tissue samples used for this study were obtained with written informed consent from all participants in accordance with the guidelines in The Declaration of Helsinki 2000. Single-nuclei extraction: Fresh tissue samples of human implantation sites were embedded in cold OCT medium and flash frozen using a dry ice-isopentane slurry. Protocol available at doi:10.17504/protocols.io.95mh846. Single-nuclei suspensions were isolated from frozen tissue sections following manufacturer's instructions. For each OCT-embedded sample, 400 μm of tissue was prepared as 50 μm cryosections, which were paused in a tube on dry ice until subsequent processing. Nuclei were released via Dounce homogenisation. Single-cell extraction: We used the previous protocol optimised for the decidual-placental interface (Vento-Tromo et al 2018, Nature). Briefly, decidual tissues were enzymatically digested in 15 ml 0.4 mg/ml collagenase V (Sigma, C-9263) solution in RPMI 1640 medium (Thermo Fisher Scientific, 21875-034)/10% FCS (Biosfera, FB-1001) at 37 °C for 45 min. The supernatant was diluted with medium and passed through a 100-μm cell sieve (Corning, 431752) and then a 40-μm cell sieve (Corning, 431750). The flow-through was centrifuged and resuspended in 5 ml of red blood cell lysis buffer (Invitrogen, 00-4300) for 10 min. Placental villi were scraped from the chorionic membrane using a scalpel and the stripped membrane was discarded. The resultant villous tissue was enzymatically digested in 70 ml 0.2% trypsin 250 (Pan Biotech P10-025100P)/0.02% EDTA (Sigma E9884) in PBS with stirring at 37 °C for 9 min. The disaggregated cell suspension was diluted with medium and passed through a 100-μm cell sieve (Corning, 431752). The undigested gelatinous tissue remnant was retrieved from the gauze and further digested with 10-15 ml collagenase V at 1.0 mg/ml (Sigma C9263) in Ham’s F12 medium/10% FBS with gentle shaking at 37 °C for 10 min. The disaggregated cell suspension was diluted with medium and passed through a 100-μm cell sieve (Corning, 431752). Cells obtained from both enzyme digests were pooled together and passed through a 100-μm cell sieve (Corning, 431752) and washed in Ham’s F12. The flow-through was centrifuged and resuspended in 5 ml of red blood cell lysis buffer (Invitrogen, 00-4300) for 10 min. For the scRNA-seq experiments, cells were loaded according to the manufacturer’s protocol for the Chromium Single Cell 3′ Kit v3.0, v3.1 and 5’ v1.0 (10X Genomics). Library preparation was carried out according to the manufacturer’s protocol to attain between 2,000 and 10,000 cells per reaction. For the snRNA-seq experiments, nuclei were loaded according to the manufacturer’s protocol for the Chromium Single Cell Multiome ATAC + Gene Expression v1.0 to attain between 2,000 and 10,000 cells per well. Library preparation was carried out according to the manufacturer’s protocol.
characteristics.characteristic: description
characteristics.text: embryo
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: H2
characteristics.characteristic: individual
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: placenta
characteristics.characteristic: organism part
characteristics.text: single-cell RNA sequencing
characteristics.characteristic: protocol
characteristics.text: maternal fetal interface
characteristics.characteristic: sampling site
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Pla_HDBR8624430
characteristics.characteristic: title