biostudies-arrayexpressXiangning DongHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15358We performed single cell RNA sequencing (scRNA-seq) of human fetal duodenal tissue samples from a single individual biological specimen at 70 days post-conception (DPC). The data set is composed of cells from diverse intestinal lineages. Lineages captured include epithelium, mesenchyme, immune, neurons, and endothelium. These data were used to specifically interrogate the development and emergence of mesenchymal lineages during human development. This data was combined with previously published data from our lab (E-MTAB-9489, E-MTAB-10187, and E-MTAB-9906) to generate an scRNA-seq database of human small intestinal development.biostudies-arrayexpressNucleic Acid Extraction - Immediately after tissue dissociation, single cell libraries were prepared on the 10x Chromium with a target of 5,000 cells. Chromium Single Cell 3’ Library & Gel Bead Kit v2, 16 rxns PN-120237 was used.Library Construction - Immediately after tissue dissociation, single cell libraries were prepared on the 10x Chromium with a target of 5,000 cells. Chromium Single Cell 3’ Library & Gel Bead Kit v2, 16 rxns PN-120237 was used.Sequencing - All single-cell RNA-sequencing was performed with an Illumina HiSeq 4000 by the University of Michigan DNA Sequencing core.Sample Collection - Use of human tissue was reviewed and approved by The University of Michigan Institutional Review Board (IRB). De-identified human fetal intestine tissue was obtained from the University of Washington Laboratory of Developmental Biology. Tissue was shipped overnight in Belzer-UW Cold Storage Solution (ThermoFisher, NC0952695) with cold packs. Cell dissociations were carried out similar to previously published methods (Miller et al., 2020). To dissociate human fetal tissue to single cells, tissue was mechanically minced into small fragments, and in a petri dish filled with ice-cold 1X HBSS (with Mg2+, Ca2+). This tissue was then transferred to a 15 mL conical tube. Dissociation enzymes and reagents from the Neural Tissue Dissociation Kit (Miltenyi, cat. no. 130-092-628) were used, and all incubation steps were carried out in a refrigerated centrifuge pre-chilled to 10ºC unless otherwise stated. All tubes and pipette tips used to handle cell suspensions were pre-washed with 1% BSA in HBSS to prevent adhesion of cells to the plastic. Tissue was treated for 15 minutes at 10ºC with Mix 1. Mix 2 was added to the digestion, and tissue was incubated for 10 minute increments at 10ºC until digestion was complete. After each 10 minute incubation, tissue was agitated using a P1000, and tissue digestion was visually assessed under a stereo microscope. This process continued until the tissue was fully digested. Cells were filtered through a 70 µm filter coated with 1% BSA in 1X HBSS, spun down at 500g for 5 minutes at 10ºC and resuspended in 500µl 1X HBSS (with Mg2+, Ca2+). 1 mL Red Blood Cell Lysis buffer (Roche cat. No 11814389001) was then added to the tube and the cell mixture was placed on a rocker for 15 minutes at 4ºC. Cells were spun down (500g for 5 minutes at 10ºC), and washed twice by suspension in 2 mLs of HBSS + 1% BSA followed by centrifugation. Cells were counted using a hemocytometer, then spun down and resuspended (if necessary) to reach a concentration of 1000 cells/µL and kept on ice. Single cell droplets were immediately prepared on the 10x Chromium according to manufacturer instructions at the University of Michigan The Advanced Genomics Core, with a target of capturing 5,000 cells. Single cell libraries were prepared using the Chromium Next GEM Single Cell 3’ Library Construction Kit (v2) according to manufacturer instructions. After single cell dissociation, cells were stained with DAPI and sorted for viability using FACS. FACS was performed on Sony Synergy Head cell sorter.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Data matrices for further analysis were generated using the CellRanger pipeline (v) under standard parameters by the University of Michigan Advanced Genomics Sequencing Core.UnknownTranscriptomicsGenomicsProteomics10x Chromium with a target of 5,000 cells. Chromium Single Cell 3’ Library & Gel Bead Kit v2NAIllumina HiSeq 400010X Genomics reagent kitRNA-seq of coding RNA from single cellsHomo sapiensMapping mesenchymal diversity in the developing human intestine and organoidsKelli Johnson, Xiangning Dong, Yu-Hwai Tsai, Angeline Wu, Sydney G. Clark, Rachel Zwick, Ian Glass, Katherine D. Walton, Ophir Klein, Jason R. SpenceXiangning DongKelli JohnsonJason SpencefalsescRNAseq of human fetal duodenum, 70 DPCWe performed single cell RNA sequencing (scRNA-seq) of human fetal duodenal tissue samples from a single individual biological specimen at 70 days post-conception (DPC). The data set is composed of cells from diverse intestinal lineages. Lineages captured include epithelium, mesenchyme, immune, neurons, and endothelium. These data were used to specifically interrogate the development and emergence of mesenchymal lineages during human development. This data was combined with previously published data from our lab (E-MTAB-9489, E-MTAB-10187, and E-MTAB-9906) to generate an scRNA-seq database of human small intestinal development.2025-07-30T00:00:00Z2025-07-14T13:32:38.471Z2025-07-14T13:32:38.471ZE-MTAB-15358ERP176756E-MTAB-9489E-MTAB-9906E-MTAB-10187EFO_0002944EFO_0004170EFO_0005684EFO_0005518EFO_0003816EFO_0004184