description: Protocols: All 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. Human embryo and fetal samples were obtained from the MRC and Wellcome-funded Human Developmental Biology Resource (HDBR43, http:// www.hdbr.org), with appropriate maternal written consent and approval from the Newcastle and North Tyneside NHS Health Authority Joint Ethics Committee (08/H0906/21+5). The HDBR is regulated by the UK Human Tissue Authority (HTA; www.hta.gov.uk) and operates in accordance with the relevant HTA Codes of Practice. We used the previous protocol optimised for gonadal dissociation and available at protocols.io. https://www.protocols.io/view/human-embryonic-gonad-dissociation-with-trypsin-ed-66fhhbn In short, tissues were cut into <1 mm segments before being transferred to a 15ml tube. Tissues were digested with Trypsin/EDTA 0.25% between 5-15 minutes at 37°C with intermittent shaking. Digested tissue was passed through a 100 µm filter, and cells collected by centrifugation (500 x g for 5 minutes at 4°C). Cells were washed with flow buffer (PBS containing 5 % (v/v) FBS and 2 mM EDTA) prior to cell counting. library For the scRNA-seq experiments, cells were loaded according to the manufacturer's protocol for the Chromium Single Cell 5′ Kit v.1.0, v1.1 and v.2 (10X Genomics) to attain between 2,000 and 10,000 cells per reaction. Library preparation was carried out according to the manufacturer's protocol. Library preparation was carried out according to the manufacturer's protocol for the Chromium Single Cell 5′ Kit v.1.0, v1.1 and v.2 (10X Genomics).
attributes.tag: ENA first public
attributes.value: 2022-05-26
attributes.tag: ENA last update
attributes.value: 2022-05-26
attributes.tag: External Id
attributes.value: SAMEA14322276
attributes.tag: INSDC center alias
attributes.value: Wellcome Sanger Institute
attributes.tag: INSDC center name
attributes.value: Wellcome Sanger Institute
attributes.tag: INSDC first public
attributes.value: 2022-05-26T08:13:58Z
attributes.tag: INSDC last update
attributes.value: 2022-05-26T08:13:58Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10551_2:FCA_GND9295209
attributes.tag: age
attributes.value: 16 gestational week
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: gonad
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: embryo
attributes.tag: disease
attributes.value: normal
attributes.tag: individual
attributes.value: F100
attributes.tag: organism part
attributes.value: gonad
attributes.tag: sample name
attributes.value: E-MTAB-10551_2:FCA_GND9295209
attributes.tag: sex
attributes.value: male