Project description:Fetal lung samples at 12–20 post conception week (pcw) from the HDBR, up to 0.5cm3 in size, were embedded in OCT and flash-frozen in dry-ice cooled isopentane. Twelve-micron cryosections were cut onto Visium slides, haematoxylin and eosin stained and imaged at 20X magnification on a Hamamatsu Nanozoomer 2.0 HT Brightfield. These were then further processed according to the 10X Genomics Visium protocol, using a permeabilization time of 18 min for 12–17 pcw samples and 24 min for 19 pcw and older samples. Images were exported as tiled tiffs for analysis. Dual-indexed libraries were prepared as in the 10X Genomics protocol, pooled at 2.25 nM and sequenced in 4 samples per Illumina Novaseq SP flow cell with read lengths of 28 bp for R1, 10 bp for i7 index, 10 bp for i5 index, 90 bp for R2.
Project description:Cancer cells display highly heterogeneous and plastic states in glioblastoma, an incurable brain tumour. However, how these malignant states arise and whether they follow defined cellular trajectories across tumours is poorly understood. Here, we generated a deep single cell and spatial multi-omic atlas of human glioblastoma that pairs transcriptomic, epigenomic and genomic profiling of 12 tumours across multiple regions. Please cite De Jong et al., bioRxiv 2025 upon use. This submission contains the Space Ranger output bundles from Visium spatial transcriptomic sequencing (10x Genomics), including paired high-resolution H&E tissue images, for each of the 97 Visium sections generated as part of the GBM-space atlas. See README_spaceranger.md. We also include integrated single cell and spatial anndata representations of all spatial transcriptomic data from our 97 Visium sections. Feature types include gene expression, cell state abundances, spatial niche abundances, and histopathological annotations for each spatial tissue location. See README_visium_anndata.md.
Project description:Individual TFs were overexpressed in fetal lung tip organoids from a doxycycline-inducible construct for 3 days, and organoids were maintained in the self-renewing (tip cell-promoting) medium throughout to rigorously assay the lineage-determining competence of the TF, followed by scRNA-seq. ASCL1, NEUROD1, and NEUROG3 were selected as key neuroendocrine regulators. We also selected the GHRL+ NE-specific RFX6 and NKX2.2, the pan-NE PROX1, and, as controls, the basal cell-specific TFs DeltaNTP63, TFAP2A, PAX9, and mNeonGreen-3xNLS.
Project description:We obtained human embryonic and fetal lungs from 5-22 pcw for scRNAseq and scATACseq analysis. To focus on epithelial differentiation and region specialization, we deeply sampled 15, 18, 20 and 22 pcw lungs and separated proximal and distal regions while leaving lungs at 5, 6, 9 and 11 pcw intact. These cell samples (except for one at 6pcw) were split and processed for both scRNAseq and scATACseq.
Project description:We obtained human embryonic and fetal lungs from 5-22 pcw for scRNAseq and scATACseq analysis. To focus on epithelial differentiation and region specialization, we deeply sampled 15, 18, 20 and 22 pcw lungs and separated proximal and distal regions while leaving lungs at 5, 6, 9 and 11 pcw intact. These cell samples (except for one at 6pcw) were split and processed for both scRNAseq and scATACseq.