Project description:The conducting airway epithelium of the rodent and human lung is made up of about equal proportions of ciliated and secretory cells. In addition, in regions where the epithelium is pseudostratfied, ~30% of the epithelium consists of undifferentiated basal cells (BCs). Evidence suggests that these BCs are multipotent stem cells that can self renew over the long term and give rise to both ciliated and secretory lineages. The goal of this project is to identify cellular and molecular mechanisms by which the basal cells normally maintain the epithelium and repair it after injury. We used Affymetrix microarray analysis to compare transcripts in tracheal epithelium before and after SO2 injury. Mice tracheal epithelium and mesenchyme were separate for RNA extraction before and 48hrs after SO2 injury. Cells from tracheas of 4 male C57Bl/6 mice were pooled for each biological experiment. The experiments were repeated three times for triplicate samples.
Project description:The conducting airway epithelium of the rodent and human lung is underlaid by mesenchymal cells that include vasculature, smooth muscle, fibroblasts and cartilage. The goal of this project is to identify cellular and molecular changes in the mesenchyme after injury to the epithelium by exposure to SO2 and which may participate in repair of the epithelium We used Affymetrix microarray analysis to compare transcripts in tracheal mesenchyme before and after SO2 injury. Mice tracheal epithelium and mesenchyme were separate for RNA extraction before and 48hrs after SO2 injury. Sample from 4 mice were pooled for each biological experiment. The experiments were repeated three times for triplicate samples.
Project description:We performed microarray analysis to assess changes in gene expression in dysplastic lesions in the mouse oesophagus versus adjacent normal epithelium. To induce tumor formation, animals were treated with the tobacco-derived carcinogen DEN (diethylnitrosamine) in drinking water 3 times a week for 56 days. Subsequently, Sorafenib was administered by ip injection as a solution at 10 mg/ml for 28 days. RNA was isolated from 9 HGD (high grade dysplasia) samples and 9 control samples of adjacent normal epithelium from the same animal (n= 9 samples from a total of 6 animals). Hybridisation was performed on a Mouse Whole Genome-6 v2 Illumina array.
Project description:Tumor Associated Calcium Signal Transducer 2 (TACSTD2) is one of the cancer-related genes whose overexpression correlates with tumor progression and invasiveness in human colorectal cancer. TACSTD2 gene encodes for a transmembrane glycoprotein TROP2, which is implicated in altered expression of epithelial-mesenchymal transition (EMT) markers and may play a role in metastasis formation. To determine how TROP2 affects aberrant tumor cell signaling, we isolated early adenoma cells from the mouse small intestine 6 weeks after disruption of the Adenomatous polyposis coli (Apc) gene, one of the first steps in the development of colorectal cancer in human. We performed expression profiling of Trop2+ and Trop2- tumor cells and, in addition, non-tumor cells of the intestinal epithelium, including predominantly differentiated cells.
Project description:Tumor Associated Calcium Signal Transducer 2 (TACSTD2) is one of the cancer-related genes whose overexpression correlates with tumor progression and invasiveness in human colorectal cancer. TACSTD2 gene encodes for a transmembrane glycoprotein TROP2, which is implicated in altered expression of epithelial-mesenchymal transition (EMT) markers and may play a role in metastasis formation. To monitor the onset of TROP2 in hyperplastic cells and its effect on aberrant signaling potentially leading to tumor growth, we isolated epithelial cells from the mouse small intestine 7 days after disruption of the Adenomatous polyposis coli (Apc) gene. Loss of APC tumor suppressor function is the most common initial step in the development of colon cancer in humans. In the mice, the epithelial hyperproliferation occurs within a few days after Apc disruption, accompanied by the formation of ectopic crypts, followed by the formation of microadenomas over time. We performed expression profiling of Trop2+ and Trop2- hyperproliferating cells and, in addition, non-proliferating cells of the intestinal epithelium, including predominantly differentiated cells.
Project description:The peptide-level analysis of proteome and secretome changes of mouse trachea cells upon denatonium treatment (in comparison to Ringer lactate solution control).
Project description:We previously identified multipotent stem cells within the lamina propria of the human olfactory mucosa, located in the nasal cavity. We also demonstrated that this cell type differentiates into neural cells and improves locomotor behavior after transplantation in a rat model of Parkinsonâs disease. Yet, next to nothing is known about their specific stemness characteristics. We therefore devised a study aiming to compare olfactory lamina propria stem cells from 4 individuals to bone marrow mesenchymal stem cells from 4 age- and gendermatched individuals. Using pangenomic microarrays and immunostaining with 34 cell surface marker antibodies, we show here that olfactory stem cells are closely related to bone marrow stem cells. However, olfactory stem cells exhibit also singular traits. By means of techniques such as proliferation assay, cDNA microarrays, RT-PCR, in vitro and in vivo differentiation, we report that, when compared to bone marrow stem cells, olfactory stem cells display i) a high proliferation rate; ii) a propensity to differentiate into osseous cells and iii) a disinclination to give rise to chondrocytes and adipocytes. Since peripheral olfactory stem cells originate from a neural crest-derived tissue and, as shown here, exhibit an increased expression of neural cellrelated genes, we propose to name them olfactory ecto-mesenchymal stem cells (OE-MSC). Further studies are now required to corroborate the therapeutic potential of OE-MSCs in animal models of bone and brain diseases. Keywords: cell type comparison Expression profiles of olfactory ectomesenchymal stem cells, bone marrow MSC, periosteal cells, neural progenitors, and synovial fibroblasts were compared against each other and with different lineages of purified hematopoietic cells from bone marrow to characterise human olfactory ectomesenchymal stem cells molecularly
Project description:Recent studies have shown that tissue macrophages (MF) arise from embryonic progenitors of the yolk sac (YS) and fetal liver and colonize the tissues before birth. Further studies have proposed that developmentally distinct tissue MF can be identified based on the differential expression of F4/80 and CD11b, but whether a characteristic transcriptional profile exists is largely unknown. Here, we established an inducible fate mapping system that facilitated the identification of A2 progenitors of the YS as source of F4/80hi but not CD11bhi MF. Large-scale transcriptional profiling of MF precursors from the YS until adulthood allowed the description of a complex MF pedigree. We further identified a distinct molecular signature of F4/80hi and CD11bhi MF and found that Irf8 was vital for MF maturation and the innate immune response. Our data provide new cellular and molecular insights into the origin and developmental pathways of tissue MF. All samples are from mouse tissue at early developmental stages (E8, E9.5, E14) and from adulthood (6 weeks old). For the early developmental time points timed matings were performed. Macrophage populations were isolated from each tissue. RNA was isolated using Arcturus PicoPure isolation kit from yolk sac, brain, liver, kidney and skin samples after FACS sorting. Three replicates per cell population were included. Wildtype and Irf8 knockout samples were analyzed.
Project description:Label free: The aim of project is to identify the proteins that may have a role in the inductive signal of mesenchymal cells. Protein identification and quantitively compare the proteins from the following samples: 1- Exosomes purified from embryonic mesenchymal cells of tooth germ after overnight culture in 2D and 3D conditions 2- Secretome of embryonic mesenchymal cells of tooth germ after overnight 3D culture. 3- Embryonic mesenchymal cells from tooth germ- fresh frozen as pellet TMT Samples 10.5 and 14.5 are exosomes derived from immortalized cell line and different stages of tooth embryonic development. These sets of samples were seeded in well plate. The embryonic samples are a pool of 6 biological replicates. The aim is to identify the different proteins between these stages as well as the cell line.
Project description:Stratification of breast cancers into subtypes are generally based on immune assays on tumor cells and/or mRNA expression of tumor cell enriched tissues. Here, we have laser microdissected tumor epithelium and tumor stroma from 24 breast cancer biopsies (12 luminal-like and 12 basal-like). We hypothesized that the stromal proteome would separate patients with breast into groups independently of the traditional epithelial based subtypes.