Stem cells and fluid flow drive cyst formation in an invertebrate excretory organ.
Ontology highlight
ABSTRACT:
SUBMITTER: Thi-Kim Vu H
PROVIDER: S-EPMC4500094 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:Liquid in a Petri dish spontaneously circulates in a radial pattern, even when the dish is at rest. These fluid flows have been observed and utilized for biological research, but their origins have not been well-studied. Here we used particle-tracking to measure velocities of radial fluid flows, which are shown to be linked to evaporation. Infrared thermal imaging was used to identify thermal gradients at the air-liquid interface and at the bottom of the dish. Two-color ratiometric fluorescence confocal imaging was used to measure thermal gradients in the vertical direction within the fluid. A finite-element model of the fluid, incorporating the measured temperature profiles, shows that buoyancy forces are sufficient to produce flows consistent with the measured particle velocity results. Such flows may arise in other dish or plate formats, and may impact biological research in positive or negative ways.
| S-EPMC4856293 | biostudies-literature
Project description:In common with most mammals, humans form only two dentitions during their lifetime. Occasionally, supernumerary teeth develop in addition to the normal complement. Odontoma represent a small group of malformations containing calcified dental tissues of both epithelial and mesenchymal origin, with varying levels of organization, including tooth-like structures. The specific cell type responsible for the induction of odontoma, which retains the capacity to re-initiate de novo tooth development in postnatal tissues, is not known. Here we demonstrate that aberrant activation of WNT signaling by expression of a non-degradable form of β-catenin specifically in SOX2-positive postnatal dental epithelial stem cells is sufficient to generate odontoma containing multiple tooth-like structures complete with all dental tissue layers. Genetic lineage-tracing confirms that odontoma form in a similar manner to normal teeth, derived from both the mutation-sustaining epithelial stem cells and adjacent mesenchymal tissues. Activation of the WNT pathway in embryonic SOX2-positive progenitors results in ectopic expression of secreted signals that promote odontogenesis throughout the oral cavity. Significantly, the inductive potential of epithelial dental stem cells is retained in postnatal tissues, and up-regulation of WNT signaling specifically in these cells is sufficient to promote generation and growth of ectopic malformations faithfully resembling human odontoma.
| S-EPMC4585991 | biostudies-other
Project description:Genomic and genetic analyses have demonstrated that many species contain multiple chemotaxis-like signal transduction cascades that likely control processes other than chemotaxis. The Che? signal transduction cascade from Rhodospirillum centenum is one such example that regulates development of dormant cysts. This Che-like cascade contains two hybrid response regulator-histidine kinases, CheA? and CheS?, and a single-domain response regulator CheY?. We demonstrate that cheS? is epistatic to cheA? and that only CheS??P can phosphorylate CheY?. We further show that CheA? derepresses cyst formation by phosphorylating a CheS? receiver domain. These results demonstrate that the flow of phosphate as defined by the paradigm E. coli chemotaxis cascade does not necessarily hold true for non-chemotactic Che-like signal transduction cascades.
| S-EPMC3868531 | biostudies-literature
Project description:BACKGROUND: There is little knowledge concerning the content and the mechanisms of filling of arachnoid cysts. The aim of this study was to compare the protein content of arachnoid cysts and cerebrospinal fluid by quantitative proteomics to increase the understanding of arachnoid cysts. METHODS: Arachnoid cyst fluid and cerebrospinal fluid from five patients were analyzed by quantitative proteomics in two separate experiments.In a label-free experiment arachnoid cyst fluid and cerebrospinal fluid samples from individual patients were trypsin digested and analyzed by Orbitrap mass spectrometry in a label-free manner followed by data analysis using the Progenesis software.In the second proteomics experiment, a patient sample pooling strategy was followed by MARS-14 immunodepletion of high abundant proteins, trypsin digestion, iTRAQ labelling, and peptide separation by mix-phase chromatography followed by Orbitrap mass spectrometry analysis. The results from these analyzes were compared to previously published mRNA microarray data obtained from arachnoid membranes. RESULTS: We quantified 348 proteins by the label-free individual patient approach and 1425 proteins in the iTRAQ experiment using a pool from five patients of arachnoid cyst fluid and cerebrospinal fluid. This is by far the largest number of arachnoid cyst fluid proteins ever identified, and the first large-scale quantitative comparison between the protein content of arachnoid cyst fluid and cerebrospinal fluid from the same patients at the same time. Consistently in both experiment, we found 22 proteins with significantly increased abundance in arachnoid cysts compared to cerebrospinal fluid and 24 proteins with significantly decreased abundance. We did not observe any molecular weight gradient over the arachnoid cyst membrane. Of the 46 proteins we identified as differentially abundant in our study, 45 were also detected from the mRNA expression level study. None of them were previously reported as differentially expressed. We did not quantify any of the proteins corresponding to gene products from the ten genes previously reported as differentially abundant between arachnoid cysts and control arachnoid membranes. CONCLUSIONS: From our experiments, the protein content of arachnoid cyst fluid and cerebrospinal fluid appears to be similar. There were, however, proteins that were significantly differentially abundant between arachnoid cyst fluid and cerebrospinal fluid. This could reflect the possibility that these proteins are affected by the filling mechanism of arachnoid cysts or are shed from the membranes into arachnoid cyst fluid. Our results do not support the proposed filling mechanisms of oncotic pressure or valves.
| S-EPMC3641952 | biostudies-literature
Project description:Background:
* Researchers are trying to learn what causes certain types of cancer to spread to other organs in the body (metastasis). Cancer tumors may produce a very small number of specific cells (cancer stem cells) that cause the tumors to grow in other organs throughout the body.
* By examining cancer tumor tissue, normal tissue, blood, bone marrow, and other body fluids, researchers may determine whether these samples contain cancer stem cells. Cancer stem cells may provide information on whether the cancer will come back or spread before other routine x-ray studies or lab tests indicate its presence.
Objectives:
* To acquire a collection of solid organ cancer stem cells for future study.
* To analyze solid organ cancer stem cells from various types of cancer on a genetic level.
* To determine if solid organ cancer stem cells are present in the blood or bone marrow.
Eligibility:
* Patients 16 years of age and older who have solid organ cancer (cancer in the liver, colon, rectum, anus, pancreas, stomach, breast, skin, muscles, fat, connective tissue, uterus, ovary, cervix, vagina, vulva, or inner lining of the abdomen) or a precancerous growth, and who are scheduled to have a biopsy or surgery to remove the cancer as part of their treatment.
Design:
* This is a prospective trial designed to procure solid organ cancer stem cells before either surgery or biopsy.
* All patients registered to this trial will undergo surgery to extirpate their cancer in the NCI
* Prior to surgery or biopsy, 8 tablespoons of blood will be drawn.
* During the surgery or biopsy, a sample of normal tissue will be removed along with the cancerous or precancerous tissue. If separate consent is given, samples of bone marrow will also be taken.
* After discharge, patients will return to the clinic for routine visits every month for the first 3 months following surgery, and then about every 3 months for 2 years, and then every 6 months for 3 years. During the visits, patients will have routine blood and imaging studies done, and researchers will take additional blood samples (about 8 tablespoons at each visit) and optional bone marrow samples (4 teaspoons every 6 months) to be used for research.
| 2073925 | ecrin-mdr-crc
Project description:Doublecortin-like kinase 1 (DCLK1)-positive pancreatic cancer stem cells develop at a precancerous stage and may contribute to the lack of efficacy of pancreatic cancer therapy. Although PanIN cells express oncogenic KRas and have an increased activity of epidermal growth factor receptor (EGFR), we demonstrate that, in DCLK1+ PanIN cells, EGFR signaling is not propagated to the nucleus. Mimicking blockage of EGFR with erlotinib in PanIN organoid culture or in p48cre;KrasG12D mice led to a significant increase in DCLK1+ PanIN cells. As a mechanism of how EGFR inhibition leads to formation of DCLK1+ cells, we identify an increase in hydrogen peroxide contributing to activation of Protein Kinase D1 (PKD1). Active PKD1 then drives stemness and abundance of DCLK1+ cells in lesions. Our data suggest a signaling mechanism that leads to the development of DCLK1+ pancreatic cancer stem cells, which can be exploited to target this population in potential therapeutic approaches.
| S-EPMC7820128 | biostudies-literature
Project description:Cytosolic fluid dynamics have been implicated in cell motility because of the hydrodynamic forces they induce and because of their influence on transport of components of the actin machinery to the leading edge. To investigate the existence and the direction of fluid flow in rapidly moving cells, we introduced inert quantum dots into the lamellipodia of fish epithelial keratocytes and analysed their distribution and motion. Our results indicate that fluid flow is directed from the cell body towards the leading edge in the cell frame of reference, at about 40% of cell speed. We propose that this forward-directed flow is driven by increased hydrostatic pressure generated at the rear of the cell by myosin contraction, and show that inhibition of myosin II activity by blebbistatin reverses the direction of fluid flow and leads to a decrease in keratocyte speed. We present a physical model for fluid pressure and flow in moving cells that quantitatively accounts for our experimental data.
| S-EPMC2867054 | biostudies-literature
Project description:Human mesenchymal stem cells (hMSCs) have the potential to differentiate into different types of mesodermal tissues. In vitro proliferation and differentiation of hMSCs are necessary for bone regeneration in tissue engineering. The present study aimed to design and develop a fluid flow mechanically-assisted cartridge device to enhance the osteogenic differentiation of hMSCs. We used the fluorescence-activated cell-sorting method to analyze the multipotent properties of hMSCs and found that the cultured cells retained their stemness potential. We also evaluated the cell viabilities of the cultured cells via water-soluble tetrazolium salt 1 (WST-1) assay under different rates of flow (0.035, 0.21, and 0.35 mL/min) and static conditions and found that the cell growth rate was approximately 12% higher in the 0.035 mL/min flow condition than the other conditions. Moreover, the cultured cells were healthy and adhered properly to the culture substrate. Enhanced mineralization and alkaline phosphatase activity were also observed under different perfusion conditions compared to the static conditions, indicating that the applied conditions play important roles in the proliferation and differentiation of hMSCs. Furthermore, we determined the expression levels of osteogenesis-related genes, including the runt-related protein 2 (Runx2), collagen type I (Col1), osteopontin (OPN), and osteocalcin (OCN), under various perfusion vis-à-vis static conditions and found that they were significantly affected by the applied conditions. Furthermore, the fluorescence intensities of OCN and OPN osteogenic gene markers were found to be enhanced in the 0.035 mL/min flow condition compared to the control, indicating that it was a suitable condition for osteogenic differentiation. Taken together, the findings of this study reveal that the developed cartridge device promotes the proliferation and differentiation of hMSCs and can potentially be used in the field of tissue engineering.
| S-EPMC8398302 | biostudies-literature
Project description:Microdroplets and microcapsules have been widely produced using microfluidic flow-focusing junctions for biomedical and chemical applications. However, the multiphase microfluidic flow at the flow-focusing junction has not been well investigated. In this study, the displacement of two (core and shell) aqueous fluids that disperse into droplets altogether in a carrier oil emulsion was investigated both numerically and experimentally. It was found that extensive displacement of the two aqueous fluids within the droplet during its formation could occur as a result of the shear effect of the carrier fluid and the capillary effect of interfacial tension. We further identified that the two mechanisms of fluid displacement can be evaluated by two dimensionless parameters. The quantitative relationship between the degree of fluid displacement and these two dimensionless parameters was determined experimentally. Finally, we demonstrated that the degree of fluid displacement could be controlled to generate hydrogel microparticles of different morphologies using planar or nonplanar flow-focusing junctions. These findings should provide useful guidance to the microfluidic production of microscale droplets or capsules for various biomedical and chemical applications.
| S-EPMC4605896 | biostudies-other
Project description:The aim of the present study was to investigate the stimulation of osteoinduced mesenchymal stem cells (MSCs) into chondrogenically predifferentiated MSCs and chondrocytes in a mechanical environment. A novel two?layer microfluidic chip was used to mimic the interstitial flow in the superficial zones of articular cartilage. The morphology, proliferation rate and the expression of collagen I, collagen II and aggrecan of chondrocytes and chondro?MSCs were investigated. The results revealed that the cells in the bottom layer were influenced by the top layer's osteoinduced MSCs and the bottom layer's shear flow. The expression of collagen I, which may signify the effect of the shear stress on the dedifferentiation change, was weakened by the stimulation of osteoinduced MSCs on the top layer. The expression of collagen II and aggrecan was increased in the fluidic environment by osteoinduced MSCs. These results indicate that osteoinduced MSCs have a significant effect on the phenotype of chondro?MSCs and chondrocytes in the fluidic microenvironment. The present study described a simple and promising way to rapidly evaluate cell responses to other cells in a fluidic environment, which may help to better promote the utilization of MSCs and chondrocytes in tissue engineering.
| S-EPMC5783459 | biostudies-literature