Prowling wolves in sheep's clothing: the search for tumor stem cells

2008 ◽  
Vol 389 (7) ◽  
Author(s):  
Ibrahim Alkatout ◽  
Dieter Kabelitz ◽  
Holger Kalthoff ◽  
Sanjay Tiwari
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Surface Protein ◽  
Tumor Initiation ◽  
Tumor Initiating Cells ◽  
Resource Guide ◽  
Therapeutic Implications ◽  
Biological Fate ◽  
Surface Protein Expression

AbstractThe importance of a subset of cells which have ‘stem like’ characteristics and are capable of tumor initiation has been reported for a range of tumors. Isolation of these tumor-initiating cells (TICs) has largely been based on differential cell surface protein expression. However, there is still much debate on the functional significance of these markers in initiating tumors, as many properties of tumor initiation are modified by cell-cell interactions. In particular, the relationship between TICs and their microenvironment is poorly understood but has therapeutic implications, as the microenvironment can maintain tumor cells in a prolonged period of quiescence. However, a major limitation in advancing our understanding of the crosstalk between TICs and their microenvironment is the lack of sensitive techniques which allow thein vivotracking and monitoring of TICs. Application of newin vivocellular and molecular imaging technologies holds much promise in uncovering the mysteries of TIC behavior at the three-dimensional level. This review will describe recent advances in our understanding of the TIC concept and how the application ofin vivoimaging techniques can advance our understanding of the biological fate of TICs. A supplementary resource guide describing TICs from different malignancies is also presented.

scholarly journals Masticatory biomechanics in the rabbit: a multi-body dynamics analysis

2014 ◽  
Vol 11 (99) ◽  
pp. 20140564 ◽  
Author(s):  
Peter J. Watson ◽  
Flora Gröning ◽  
Neil Curtis ◽  
Laura C. Fitton ◽  
Anthony Herrel ◽  
...  
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Muscle Fibres ◽  
Micro Computed Tomography ◽  
Body Dynamics ◽  
Masticatory System ◽  
Muscle Groups ◽  
Muscle Activations ◽  
Multi Body

Multi-body dynamics is a powerful engineering tool which is becoming increasingly popular for the simulation and analysis of skull biomechanics. This paper presents the first application of multi-body dynamics to analyse the biomechanics of the rabbit skull. A model has been constructed through the combination of manual dissection and three-dimensional imaging techniques (magnetic resonance imaging and micro-computed tomography). Individual muscles are represented with multiple layers, thus more accurately modelling muscle fibres with complex lines of action. Model validity was sought through comparing experimentally measured maximum incisor bite forces with those predicted by the model. Simulations of molar biting highlighted the ability of the masticatory system to alter recruitment of two muscle groups, in order to generate shearing or crushing movements. Molar shearing is capable of processing a food bolus in all three orthogonal directions, whereas molar crushing and incisor biting are predominately directed vertically. Simulations also show that the masticatory system is adapted to process foods through several cycles with low muscle activations, presumably in order to prevent rapidly fatiguing fast fibres during repeated chewing cycles. Our study demonstrates the usefulness of a validated multi-body dynamics model for investigating feeding biomechanics in the rabbit, and shows the potential for complementing and eventually reducing in vivo experiments.

scholarly journals Phenotypic and functional characterization of synovial fluid-derived fibroblast-like synoviocytes in rheumatoid arthritis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ditte Køster ◽  
Johanne Hovgaard Egedal ◽  
Søren Lomholt ◽  
Malene Hvid ◽  
Martin R. Jakobsen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Synovial Fluid ◽  
Mononuclear Cells ◽  
Functional Characterization ◽  
Surface Protein ◽  
Tissue Destruction ◽  
Peripheral Blood Mononuclear ◽  
Surface Protein Expression ◽  
Fibroblast Like Synoviocytes

AbstractFibroblast-like synoviocytes (FLS) play an important pathological role in persistent inflammatory joint diseases such as rheumatoid arthritis (RA). These cells have primarily been characterized in the RA synovial membrane. Here we aim to phenotypically and functionally characterize cultured synovial fluid-derived FLS (sfRA-FLS). Paired peripheral blood mononuclear cells (PBMC) and sfRA-FLS from patients with RA were obtained and monocultures of sfRA-FLS and autologous co-cultures of sfRA-FLS and PBMC were established. The in situ activated sfRA-FLS were CD34-, CD45-, Podoplanin+, Thymocyte differentiation antigen-1+. SfRA-FLS expressed uniform levels of NFкB-related pathway proteins and secreted several pro-inflammatory cytokines dominated by IL-6 and MCP-1. In a co-culture model with autologous PBMC, the ICAM-1 and HLA-DR expression on sfRA-FLS and secretion of IL-1β, IL-6, and MCP-1 increased. In vivo, human sfRA-FLS were cartilage invasive both at ipsilateral and contralateral implantation site. We conclude that, sfRA-FLS closely resemble the pathological sublining layer FLS subset in terms of surface protein expression, cytokine production and leukocyte cross-talk potential. Further, sfRA-FLS are comparable to tissue-derived FLS in their capabilities to invade cartilage at implantation sites but also spread tissue destruction to a distant site. Collectively, sfRA-FLS can serve as a an easy-to-obtain source of pathological sublining FLS in RA.

scholarly journals Proteins and Molecular Pathways Relevant for the Malignant Properties of Tumor-Initiating Pancreatic Cancer Cells

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1397
Author(s):  
Lisa Samonig ◽  
Andrea Loipetzberger ◽  
Constantin Blöchl ◽  
Marc Rurik ◽  
Oliver Kohlbacher ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Function Analysis ◽  
Critical Role ◽  
Three Dimensional ◽  
Small Subset ◽  
Loss Of Function ◽  
Tumor Initiating Cells ◽  
Differential Proteomics ◽  
Pancreatic Cancer Cells

Cancer stem cells (CSCs), a small subset of the tumor bulk with highly malignant properties, are deemed responsible for tumor initiation, growth, metastasis, and relapse. In order to reveal molecular markers and determinants of their tumor-initiating properties, we enriched rare stem-like pancreatic tumor-initiating cells (TICs) by harnessing their clonogenic growth capacity in three-dimensional multicellular spheroid cultures. We compared pancreatic TICs isolated from three-dimensional tumor spheroid cultures with nontumor-initiating cells (non-TICs) enriched in planar cultures. Employing differential proteomics (PTX), we identified more than 400 proteins with significantly different expression in pancreatic TICs and the non-TIC population. By combining the unbiased PTX with mRNA expression analysis and literature-based predictions of pro-malignant functions, we nominated the two calcium-binding proteins S100A8 (MRP8) and S100A9 (MRP14) as well as galactin-3-binding protein LGALS3BP (MAC-2-BP) as putative determinants of pancreatic TICs. In silico pathway analysis followed by candidate-based RNA interference mediated loss-of-function analysis revealed a critical role of S100A8, S100A9, and LGALS3BP as molecular determinants of TIC proliferation, migration, and in vivo tumor growth. Our study highlights the power of combining unbiased proteomics with focused gene expression and functional analyses for the identification of novel key regulators of TICs, an approach that warrants further application to identify proteins and pathways amenable to drug targeting.

scholarly journals BIOM-61. FUNCTIONAL DIAGNOSTIC TESTING OF LIVE-CELL DRUG RESPONSE USING 3D PATIENT DERIVED GLIOBLASTOMA SPHEROIDS ON THE INCUCYTE PLATFORM

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii15-ii15
Author(s):  
Seth Malinowski ◽  
Mehdi Touat ◽  
Max Stockslager ◽  
Ross Giglio ◽  
Jack Geduldig ◽  
...  
Keyword(s):  
Treatment Response ◽  
Drug Response ◽  
Imaging Techniques ◽  
Diagnostic Testing ◽  
Three Dimensional ◽  
Live Cell ◽  
Live Cells ◽  
Cellular Interactions ◽  
Response Score

Abstract Three-dimensional patient derived cultures hold great potential for use as personalized functional diagnostics, enabling more accurate preclinical evaluations of drug treatments compared to conventional cell lines. Optical imaging of live cells allows for continuous, time lapsed measurements, and can provide drug response data based on rich phenotypic changes of cell cultures. However, current imaging techniques based on 2D microscopy evaluation aren’t readily adaptable to evaluate the drug response of intact spheroids, which may better represent the in vivo environment and retain critical cellular interactions within the tumor microenvironment. Using the IncuCyte live cell imaging platform, we successfully imaged a large cohort (n = 77) of patient derived glioblastoma spheroid cultures and evaluated whether changes in sphere volume could be used as a direct measure of treatment response. Improving on the default Incucyte analysis software, we developed an R data processing pipeline better suited for spheroid measurements, which quantified the heterogeneity in GBM baseline spheroid growth, and calculated a drug response score based on spheroid changes in response to DNA damaging agents (TMZ as an example). Compared to conventional viability measurements, this novel 3D drug response score was found to accurately identify both drug sensitive and resistant spheroids and showed robust concordance with genomic biomarkers of response (NGS and MGMT promoter methylation) and patient outcomes. Additionally, we coupled the 3D drug score with known genetic data to explore other key pathways and genes involved in TMZ response. We provide here novel analysis methods and public code (Github) to advance the use of IncuCyte spheroid measurements, and deconvolute 3D spheroid drug response into a quantifiable statistic. These methods are adaptable to freshly isolated patient cells for rapid evaluation of treatment response in GBM patients while remaining widely applicable to other cancers such as pancreatic, colon, and non-cancer organoids/spheroids with 3D growth.

scholarly journals High speed rotary system for catheter based 3-D imaging with optical coherence tomography (OCT)

2021 ◽  
Author(s):  
Antonio Mauro
Keyword(s):  
Optical Coherence Tomography ◽  
High Speed ◽  
Imaging Techniques ◽  
Three Dimensional ◽  
Biological Tissues ◽  
Optical Coherence ◽  
X Rays ◽  
Sound Waves ◽  
Acquisition Rate

Optical Coherence Tomography (OCT) is an addition to the other tomographic imaging techniques of x-ray computed tomography, magnetic resonance imaging, and ultrasound imaging. OCT uses optical reflections of biological tissues as opposed to x-rays, RF fields, and sound waves to obtain images. A rotary and pullback system has been developed for use with OCT. The system was developed to facilitate the three dimensional imaging of various lumens in humans and animals. The system is capable of rotating at a rate of 200 Hz. At this rate the rotary system will allow for a frame acquisition rate of 200 fps which is significantly higher than the highest published acquisition rate to date of 108 fps. The probes used with the system were modeled after the Intravascular Ultrasound (IVUS) miniature torque cable design. The probes can be sealed and sterilized between subjects without being damaged; unlike the single use IVUS probes. The rotary system was used to image the outer ear of a mouse in vivo. A lateral slice from the resulting three dimensional image was compared to the general histology of a mouse ear. The image compared well to the general anatomy as found on the histology.

scholarly journals The 3D-atlas builder: a dynamic and expandable 3D-tool for monitoring the changes in the neuronal differentiation domain during hindbrain morphogenesis

2021 ◽  
Author(s):  
Matthias Blanc ◽  
Frederic Udina ◽  
Cristina Pujades
Keyword(s):  
Neuronal Differentiation ◽  
Imaging Techniques ◽  
Three Dimensional ◽  
Temporal Data ◽  
Proof Of Concept ◽  
Experimental Proof ◽  
Atlas Construction ◽  
Dynamic Growth ◽  
Temporal Registration

Reconstruction of prototypic three-dimensional (3D) atlases at the scale of whole tissues or organs requires specific methods to be developed. We have established a protocol and provide experimental proof for building a digital 3D-atlas (here, for zebrafish hindbrain) that integrates spatial and temporal data for neuronal differentiation and brain morphogenesis, through a combination of in vivo imaging techniques paired with image analyses and segmentation tools. First, we generated a reference 3D hindbrain from several imaged specimens and segmented them using a trainable tool; these were aligned using rigid registration, revealing distribution of neuronal differentiation patterns along the axes. Second, we quantified the dynamic growth of the neuronal differentiation domain vs. the progenitor domain in the whole hindbrain. Third, we used in vivo Kaede-photoconversion experiments to generate a temporal heatmap of the neuronal growth in the whole hindbrain, revealing the spatiotemporal dynamics of neuronal differentiation upon morphogenesis. Last, as proof-of-concept, we assessed the birthdate order of GABAergic-neurons using our temporal registration map. As this protocol uses open-access tools and algorithms, it can be shared for standardized and accessible tissue-wide cell population atlas construction.

scholarly journals The lymphoid-associated interleukin 7 receptor (IL-7R) regulates tissue resident macrophage development

2019 ◽  
Author(s):  
Gabriel A. Leung ◽  
Taylor Cool ◽  
Clint H. Valencia ◽  
Atesh Worthington ◽  
Anna E. Beaudin ◽  
...  
Keyword(s):  
Fetal Liver ◽  
Surface Protein ◽  
Lineage Tracing ◽  
Late Gestation ◽  
Dynamic Regulation ◽  
Resident Macrophage ◽  
Interleukin 7 ◽  
Surface Protein Expression

AbstractThe discovery of a fetal origin for tissue-resident macrophages (trMacs) has inspired an intense search for the mechanisms underlying their development. Here, we performed in vivo lineage tracing of cells with an expression history of IL-7Rα, a marker exclusively associated with the lymphoid lineage in adult hematopoiesis. Surprisingly, we found that IL7R-Cre labeled fetal-derived, adult trMacs. Labeling was almost complete in some tissues and partial in other organs. The putative progenitors of trMacs, yolk sac (YS) erythromyeloid progenitors (EMPs), did not express IL-7R, and YS hematopoiesis was unperturbed in IL-7R-deficient mice. In contrast, tracking of IL-7Rα message levels, surface protein expression, and IL7R-Cre-mediated labeling across fetal development revealed dynamic regulation of IL-7Rα mRNA expression and rapid upregulation of IL-7Rα surface protein upon transition from monocyte to macrophage within fetal tissues. Fetal liver monocyte differentiation in vitro produced IL-7R+ macrophages, supporting a direct progenitor-progeny relationship. Additionally, blockade of IL-7R function during late gestation specifically impaired the establishment of fetal-derived tissue macrophages in vivo. These data provide evidence for a distinct function of IL-7Rα in fetal myelopoiesis and identify IL-7R as a novel regulator of tissue-resident macrophage development.

scholarly journals A MATLAB-based program for three-dimensional quantitative analysis of micronuclei reveals that neuroinflammation induces micronuclei formation in the brain

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sarasa Yano ◽  
Kaito Akiyama ◽  
Rio Tsuchiya ◽  
Hikari Kubotani ◽  
Tomoki Chiba ◽  
...  
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Innate Immune ◽  
The Other ◽  
Primary Neurons ◽  
Immune Response Pathway ◽  
The Brain ◽  
Quantitative Procedure

AbstractThe micronucleus is known to be a biomarker for genomic instability, which is a hallmark of tumors and aging. Normally, micronuclei are produced by segregation errors and mechanical stresses arising from dividing or migrating cells, leading to activation of the innate immune response pathway. Although micronuclei often emerge in damaged tissues, the quantitative procedure for analyzing micronuclei accurately has been problematic. Here, we introduce a novel MATLAB-based program for quantifying micronuclei (CAMDi: calculating automatic micronuclei distinction) in vitro and in vivo. CAMDi is adaptable to various experimental imaging techniques and is useful for obtaining reproducible data. CAMDi enables us to measure the accurate size of micronuclei from the three-dimensional images. Using CAMDi, we revealed a novel link between the emergence of micronuclei and neuroinflammation. We found that inflammatory stimulation does not increase the number of micronuclei in primary neurons. On the other hand, the administration of lipopolysaccharide into mice slightly increases micronuclei formation in neurons of the hippocampus region. These findings demonstrate that neuronal micronuclei formations are induced by an inflammatory response in a non-cell-autonomous manner. We provide a novel tool, CAMDi, to quantify micronuclei and demonstrate that neuronal micronuclei are produced not only by the cell-autonomous process but also by the intercellular communication associated with neuroinflammation in vivo.

scholarly journals N-Acetylaspartate as an in vivo Marker of Neuronal Viability in Kainate-Induced Status Epilepticus: 1H Magnetic Resonance Spectroscopic Imaging

1994 ◽  
Vol 14 (3) ◽  
pp. 373-382 ◽  
Author(s):  
Toshihiko Ebisu ◽  
William D. Rooney ◽  
Steven H. Graham ◽  
Michael W. Weiner ◽  
Andrew A. Maudsley
Keyword(s):  
Status Epilepticus ◽  
Magnetic Resonance ◽  
Imaging Techniques ◽  
Three Dimensional ◽  
Neuronal Loss ◽  
Spectroscopic Imaging ◽  
Neuronal Injury ◽  
Magnetic Resonance Spectroscopic Imaging ◽  
Neuronal Viability

N-acetylaspartate (NAA) has been proposed as a marker of neuronal density. Therefore, regional measurement of NAA by magnetic resonance spectroscopic imaging (MRSI) may provide a sensitive method for detection of selective neuronal loss, in contrast to conventional imaging techniques such as magnetic resonance imaging (MRI). To test this hypothesis, we produced selective neuronal injury by kainate-induced status epilepticus. Three days later three-dimensional 1H-MRSI was obtained and compared with conventional T2-weighted MRI and histological findings in normal and kainatetreated rats. Reduction of NAA determined by MRSI in piriform cortex, amygdala, and hippocampus correlated well with neuronal injury determined from histology. Changes of NAA, without any MRI changes in hippocampus, indicated greater sensitivity of MRSI for detection of neuronal injury. These results are consistent with the hypothesis that reduction of NAA measured by MRSI may be a sensitive marker of neuronal injury in vivo in a variety of disease states.

scholarly journals Screening of Additive Manufactured Scaffolds Designs for Triple Negative Breast Cancer 3D Cell Culture and Stem-Like Expansion

2018 ◽  
Vol 19 (10) ◽  
pp. 3148 ◽  
Author(s):  
Emma Polonio-Alcalá ◽  
Marc Rabionet ◽  
Antonio Guerra ◽  
Marc Yeste ◽  
Joaquim Ciurana ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Porous Scaffold ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Poly Lactic Acid ◽  
Tumor Initiating Cells ◽  
Fused Filament Fabrication ◽  
Open Issue

Breast cancer stem cells (BCSCs) are tumor-initiating cells responsible for metastasis and tumor reappearance, but their research is limited by the impossibility to cultivate them in a monolayer culture. Scaffolds are three-dimensional (3D) cell culture systems which avoid problems related with culturing BCSC. However, a standardized scaffold for enhancing a BCSC population is still an open issue. The main aim of this study is to establish a suitable poly (lactic acid) (PLA) scaffold which will produce BCSC enrichment, thus allowing them to be studied. Different 3D printing parameters were analyzed using Taguchi experimental design methods. Several PLA scaffold architectures were manufactured using a Fused Filament Fabrication (FFF) 3D printer. They were then evaluated by cell proliferation assay and the configurations with the highest growth rates were subjected to BCSC quantification by ALDH activity. The design SS1 (0.2 mm layer height, 70% infill density, Zigzag infill pattern, 45° infill direction, and 100% flow) obtained the highest proliferation rate and was capable of enhancing a ALDH+ cell population compared to 2D cell culture. In conclusion, the data obtained endorse the PLA porous scaffold as useful for culturing breast cancer cells in a microenvironment similar to in vivo and increasing the numbers of BCSCs.

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