Project description:The intestine is composed of an epithelial layer, containing rapidly proliferating cells that mature into two distinct anatomic regions, the small and the large intestine. Although previous studies have identified stem cells as the cell-of-origin for the whole intestine, no studies have compared stem cells derived from the small and large intestine. Here, we report intrinsic differences between these two populations of cells. Primary epithelial cells isolated from human fetal small and large intestine and expanded with Wnt agonist, R-spondin 2, displayed differential expression of stem cell markers and separate hierarchical clustering of gene expression involved in differentiation, proliferation and disease pathways. Using a three-dimensional in vitro differentiation assay, single cells derived from small and large intestine formed distinct organoid architecture with cellular hierarchy similar to that found in primary tissue. Our characterization of human fetal intestinal stem cells defies the classical definition proposed by most where small and large intestine are repopulated by an identical epithelial stem cell and raises the question of the importance of intrinsic and extrinsic cues in the development of intestinal diseases. 12 samples were analyzed. They consisted of human fetal small and large intestine (SI; n=6 and LI; n=6) stem cells, expanded with Wnt agonist and R-spondin 2. Differential expression of genes in epithelial cells from both the large and small intestine were observed.
Project description:Here, we performed single cell RNA sequencing (scRNA-seq) of 1 (one) human fetal lung tissue specimen (18.9 week) and 2 (two) human fetal intestine specimens (12.1 and 18.9 week) (total of 3 (three) independent biological specimens). The data set is composed of approximately 5,000 lung cells and 7,500 intestine cells. Lineages captured across both tissues include but are not limited to epithelium, stroma, immune, neurons and endothelium.
Project description:The intestine is composed of an epithelial layer, containing rapidly proliferating cells that mature into two distinct anatomic regions, the small and the large intestine. Although previous studies have identified stem cells as the cell-of-origin for the whole intestine, no studies have compared stem cells derived from the small and large intestine. Here, we report intrinsic differences between these two populations of cells. Primary epithelial cells isolated from human fetal small and large intestine and expanded with Wnt agonist, R-spondin 2, displayed differential expression of stem cell markers and separate hierarchical clustering of gene expression involved in differentiation, proliferation and disease pathways. Using a three-dimensional in vitro differentiation assay, single cells derived from small and large intestine formed distinct organoid architecture with cellular hierarchy similar to that found in primary tissue. Our characterization of human fetal intestinal stem cells defies the classical definition proposed by most where small and large intestine are repopulated by an identical epithelial stem cell and raises the question of the importance of intrinsic and extrinsic cues in the development of intestinal diseases.
Project description:Here, using a modified STRT-seq protocol, we analyzed the large intestine (LI) from two adults.In detail, the total 1891 single cells were picked from two adults LI. After the same rigorous filtration as fetal, 1463 individual cells were used for subsequent analysis. We identified 18 cell types including all main cell types existed in intestine such as enterocyte, goblet cell, enteroendocrine, Paneth cell, stem cell and immune cell. Take advantage of single cell RNA sequencing, we found that there were subtypes existed in some cell types. Subsequently, combined with our previous fetal data, we investigate the transcriptomic similarity and difference between human from fetal to adult. The transcriptome feature and development process from fetal to adult were revealed for the first time at single-cell resolution.
Project description:Microarray analyses of laser microdissected murine intestinal epithelial cells under the control of maternal inflammation at 17.5 days post conception demonstrates that maternal inflammation differentially regulates 2174 (iWT) and 3345 (iARE) genes in fetal tissue, but these transcriptional profiles were overwritten in the postnatal environment dominated by tissue inflammation at 8 weeks postnatal. We isolated 50 ng total RNA out of laser microdissected fetal and adult intestinal epithelial cells from the distal ileum and performed microarrays to address whether maternal inflammation programs the fetal intestine towards TNF-driven pathology
Project description:Committed progenitors with innate lymphoid cell (ILC) developmental potential are present in the fetus and bone marrow (BM). However, how fetal and BM hematopoiesis temporally and spatially contributes to ILC pools remains unclear. Here, we elucidated the distinct origins and developmental pathways of extramedullary and intramedullary ILCs during ontogeny. ILC-restricted hematopoiesis is initiated in the fetal liver (FL) and then FL-derived PD-1+ ILC progenitors (ILCPs) seed fetal lung and intestine. Organ niches determine the commitment of ILCPs to downstream precursors, including bipotent ILC1-ILC3 precursors (ILC1/3Ps) that are preferentially residing in the liver and intestine and ILC2 precursors (ILC2Ps) that show predominance in the lung. These precursors persist in adulthood and contribute to local ILC pools via BM-independent manner. In contrast, intramedullary ILC2Ps and ILC2s rely on BM hematopoiesis. Thus, our study highlights extramedullary and intramedullary ILCs with different origins, providing a comprehensive framework for developmental dynamics of ILCs.