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| Author | : Kathryn Emily Fry |
| Publisher | : |
| Total Pages | : |
| Release | : 2010 |
| Genre | : |
| ISBN | : |
| Author | : Roy Bicknell |
| Publisher | : Cambridge University Press |
| Total Pages | : 156 |
| Release | : 1996-09-28 |
| Genre | : Science |
| ISBN | : 9780521559904 |
The aim of the Handbooks in Practical Animal Cell Biology is to provide practical workbooks for those involved in primary cell culture. Each volume addresses a different cell lineage, and contains an introductory section followed by individual chapters on the culture of specific differentiated cell types. The authors of each chapter are leading researchers in their fields and use their first-hand experience to present reliable techniques in a clear and thorough manner. Endothelial Cell Culture contains chapters on endothelial cells derived from 1) lung, 2) bone marrow, 3) brain, 4) mammary glands, 5) skin, 6) adipose tissue, 7) female reproductive system, and 8) synovium.
| Author | : Doris Kogelbauer |
| Publisher | : |
| Total Pages | : 99 |
| Release | : 2014 |
| Genre | : |
| ISBN | : |
The aim of this study was to describe and impart information about the design as well as the development of an all-human in-vitro blood-brain barrier model. This in-vitro model might serve as a helpful tool for determining the permeability of the blood-brain barrier, whereby it also might decrease the necessity of in-vivo measurements in further consequence. In detail, the result of this project has been a co-cultured model, consisting of human cerebellar astrocytes (SC-1810) on the basal and human endothelial cells (hCMEC/D3) on the apical side of the model. These cells have been seeded on a membrane, which separates the basal from the apical compartment. The sum of these compartments is called two-dimensional, static Transwell model, whose integrity was determined with a Volt-Ohm Meter. The beginning section outlines basic information about the manner of functioning of the human blood-brain barrier, followed by the second main chapter, which contains details regarding those materials, which have been used for the experiments that are described in the subsequent chapter. Moreover, important cell-handling processes as well as three different cell counting methods are outlined. In the last and main chapter, one way of setting up the co-culture model is delineated, as well as the evaluation of the transendothelial electrical resistance (TEER), which also allows a conclusion regarding the integrity of the model. The described experimental set up for creating an in-vitro Transwell model might be tighten up, but basically works fine. The necessary information was obtained from intensive literature research as well as from qualitative content analysis. But primarily, this project was an empirical work, which was documented by many photographs to support the concepts.*****The aim of this study was to describe and impart information about the design as well as the development of an all-human in-vitro blood-brain barrier model. This in-vitro model might serve as a helpful tool for determining the permeability of the blood-brain barrier, whereby it also might decrease the necessity of in-vivo measurements in further consequence. In detail, the result of this project has been a co-cultured model, consisting of human cerebellar astrocytes (SC-1810) on the basal and human endothelial cells (hCMEC/D3) on the apical side of the model. These cells have been seeded on a membrane, which separates the basal from the apical compartment. The sum of these compartments is called two-dimensional, static Transwell model, whose integrity was determined with a Volt-Ohm Meter. The beginning section outlines basic information about the manner of functioning of the human blood-brain barrier, followed by the second main chapter, which contains details regarding those materials, which have been used for the experiments that are described in the subsequent chapter. Moreover, important cell-handling processes as well as three different cell counting methods are outlined. In the last and main chapter, one way of setting up the co-culture model is delineated, as well as the evaluation of the transendothelial electrical resistance (TEER), which also allows a conclusion regarding the integrity of the model. The described experimental set up for creating an in-vitro Transwell model might be tighten up, but basically works fine. The necessary information was obtained from intensive literature research as well as from qualitative content analysis. But primarily, this project was an empirical work, which was documented by many photographs to support the concepts.
| Author | : |
| Publisher | : |
| Total Pages | : |
| Release | : 2015 |
| Genre | : |
| ISBN | : |
Abstract: Metastasis is the leading cause of most cancer deaths, as opposed to dysregulated cell growth of the primary tumor. Molecular mechanisms of metastasis have been studied for decades and the findings have evolved our understanding of the progression of malignancy. However, most of the molecular mechanisms fail to address the causes of cancer and its evolutionary origin, demonstrating an inability to find a solution for complete cure of cancer. After being a neglected area of tumor biology for quite some time, recently several studies have focused on the impact of the tumor microenvironment on cancer growth. The importance of the tumor microenvironment is gradually gaining attention, particularly from the perspective of biophysics. In vitro three-dimensional (3-D) metastatic models are an indispensable platform for investigating the tumor microenvironment, as they mimic the in vivo tumor tissue. In 3-D metastatic in vitro models, static factors such as the mechanical properties, biochemical factors, as well as dynamic factors such as cell–cell, cell–ECM interactions, and fluid shear stress can be studied quantitatively. With increasing focus on basic cancer research and drug development, the in vitro 3-D models offer unique advantages in fundamental and clinical biomedical studies.
| Author | : |
| Publisher | : Newnes |
| Total Pages | : 5304 |
| Release | : 2011-08-26 |
| Genre | : Science |
| ISBN | : 0080885047 |
The second edition of Comprehensive Biotechnology, Six Volume Set continues the tradition of the first inclusive work on this dynamic field with up-to-date and essential entries on the principles and practice of biotechnology. The integration of the latest relevant science and industry practice with fundamental biotechnology concepts is presented with entries from internationally recognized world leaders in their given fields. With two volumes covering basic fundamentals, and four volumes of applications, from environmental biotechnology and safety to medical biotechnology and healthcare, this work serves the needs of newcomers as well as established experts combining the latest relevant science and industry practice in a manageable format. It is a multi-authored work, written by experts and vetted by a prestigious advisory board and group of volume editors who are biotechnology innovators and educators with international influence. All six volumes are published at the same time, not as a series; this is not a conventional encyclopedia but a symbiotic integration of brief articles on established topics and longer chapters on new emerging areas. Hyperlinks provide sources of extensive additional related information; material authored and edited by world-renown experts in all aspects of the broad multidisciplinary field of biotechnology Scope and nature of the work are vetted by a prestigious International Advisory Board including three Nobel laureates Each article carries a glossary and a professional summary of the authors indicating their appropriate credentials An extensive index for the entire publication gives a complete list of the many topics treated in the increasingly expanding field
| Author | : Jennifer L. Mantle |
| Publisher | : |
| Total Pages | : 179 |
| Release | : 2018 |
| Genre | : |
| ISBN | : 9780355758924 |
There are currently no FDA-approved therapeutics that can slow, halt, or prevent Alzheimer’s disease (AD) and only five approved drugs that treat the cognitive symptoms associated with AD. One of the key challenges with treating neurological diseases such as AD is delivery of systemically-administered therapeutics in the blood across the blood-brain barrier (BBB) into the brain. The BBB, composed of the endothelial cells that line cerebral capillaries, tightly regulates transport of molecules between the blood and the brain parenchyma and in doing so, severely limits the transport of therapeutics for neurological disease. Immunotherapies are an attractive class of therapeutic for AD due to their high target specificity and affinity however they generally exhibit notoriously low brain transport. Furthermore, while many immunotherapy drug candidates have shown efficacy in preclinical animal models, none have demonstrated disease-modifying effects in human clinical trials; studying transport of therapeutics in vivo in humans is challenging. Therefore, the goal of this research is to develop a human cell-based in vitro BBB model and to apply the model to study transport of therapeutics in AD. ☐ An ideal BBB model is made from human brain microvascular endothelial cells (BMECs), forms a tight barrier with in vivo-like transport restriction, and can be modified to mimic normal or pathological states. In this work, we differentiate human induced pluripotent stem cells into BMECs as the basis for the in vitro model which are capable of physiologically-relevant barrier performance. The model was characterized by measuring transendothelial electrical resistance (TEER), small molecule permeability, expression of BMEC-specific proteins and directional transport of a known substrate. We evaluated the permeabilities of several known small molecule drugs that can serve as benchmarks for the evaluation of new therapeutics, and validated the benchmarking system with the FDA approved AD drugs. We established a relationship between TEER and brain permeability of two different classes of drugs, suggesting fundamental differences between how small and large molecule therapeutics are transported. ☐ While studying transport of therapeutics, it is also important to consider the effects of pathological states on the BBB. AD is often accompanied by increases in plasma-derived proteins found in the brain and changes to expression or activity of transport proteins. Furthermore, molecular transport can be affected by secondary insults such as inflammation. The effects of pathological states on specific features of the BBB as well as the molecular mechanisms of immunotherapeutic transport are poorly understood. We employed a neuroinflammation model and observed impaired barrier function as measured by a decrease in barrier tightness and an increase in antibody transport. This response is partially mitigated by the presence of astrocytes. These results suggest that a breakdown in trancellular transport precedes any increase in paracellular permeability in disease and provide a link neuroinflammation and specific aspects of BBB breakdown. The model was lastly used to gain fundamental insights into the transport behavior of immunotherapies through the use of inhibitors and probes of different endocytic routes in normal, neuroinflammation and AD models. IgG transport is a saturable process and different endocytic pathways are likely responsible for IgG uptake in normal and pathological conditions. ☐ Models of the cells that comprise and surround the BBB can facilitate a more thorough understanding of disease progression, help identify new therapeutic targets, and can advance the development of new therapeutics for neurodegenerative diseases capable of reaching targets in the brain. These findings offer critical insights into the direct effects of pathological states on barrier function and demonstrate that this in vitro model can be applied to study the transport of different classes of therapeutics from the blood to the brain. Furthermore, these efforts provide a basis for future studies of transport of therapeutics at the BBB in disease, and this approach can be extended to the study of other neurological diseases and classes of therapeutics.
| Author | : Rahul Jandial |
| Publisher | : CRC Press |
| Total Pages | : 312 |
| Release | : 2013-08-08 |
| Genre | : Medical |
| ISBN | : 9781587066597 |
Most cancer deaths are a result of metastasis. The spread of a primary tumor to colonize neighboring and distant organs is the relentless endgame that defines the neoplastic process. Patients who have been diagnosed with cancer are treated to prevent both the recurrence of the tumor at the site of origin and metastasis that would re-stage them as advanced stage IV cancer. Historically and still with some types of cancer, stage IV is perceived by patients as “terminal.” Fortunately, recent molecular therapies have extended the lives of patients with advanced cancer and reassuringly people living with metastatic disease increasingly visit our clinics. What is the path forward? Given that the consilience of science and medicine is a dynamic art from which therapies arise, it would be misguided to consider any single work adequate at capturing the horizon for research. So with humility we constructed this text as primer for scientists. It begins with a broad introduction to the clinical management of common cancers. This is intended to serve as a foundation for investigators to consider when developing basic science hypotheses. Unquestionably, medical and surgical care of cancer patients reveals biology and dictates how novel therapeutics will ultimately be evaluated in clinical trials. The second section of this text offers provocative and evolving insights that underscore the breadth of science involved in the elucidation of cancer metastasis biology. The text concludes with information that integrates scientific and clinical foundations to highlight translational research. This book serves as a framework for scientists to conceptualize clinical and translational knowledge on the complexity of disease that is metastatic cancer.
| Author | : Cynthia Hajal |
| Publisher | : |
| Total Pages | : 114 |
| Release | : 2021 |
| Genre | : |
| ISBN | : |
Cancers of the brain tend to be among the most fatal, due to their rapid rates of growth and the difficulty in transporting therapeutics across the blood-brain barrier (BBB), one of the tightest vascular barrier in humans1. High-grade glioma, the most common type of primary brain cancer, has one of the worst prognoses of all cancers with a five-year survival rate of ~2.4%2−4. In addition, an estimated 20% of all cancer patients develop metastatic tumors in the brain5. Highly lethal, these stem from circulating tumor cells in brain capillaries that transmigrate into the parenchyma despite the presence of highly-regulated transport mechanisms at the BBB6−8. The lack of physiologically relevant in vitro human BBB models as well as the challenges involved in translating results from animal experiments to the clinic have significantly hindered progress in improving patient prognoses9−11. A better understanding of the mechanisms of tumor progression at the brain in a microvascular human brain-on-a-chip model that allows for high spatio-temporal resolution imaging is critical to developing new therapeutic strategies that address tumor extravasation across the BBB and glioma-BBB interactions. In this thesis, we develop an in vitro microvascular model of the human BBB in a microfluidic chip to assess the cellular and molecular interactions between cancer cells and brain stromal cells. The selfassembled BBB vascular networks are generated with induced pluripotent stem cell-derived endothelial cells, primary brain pericytes, and astrocytes. The addition of brain stromal cells resulted in improved barrier function and decreased vessel permeabilities, comparable to in vivo measurements. In addition, the engineered model has the capability to recapitulate the early steps of the metastatic cascade at the brain and primary tumor progression and interaction with the BBB in real-time via confocal microscopy. The BBB microvascular assay is then employed to obtain biological insights into the roles of brain stromal cells in the extravasation of cancer cells from various primary sites. Particularly, astrocytes are identified to play a major role in tumor transmigration through their secretion of CCL2. This chemokine is internalized by CCR2-expressing tumor cells and promotes their extravasation via both chemotaxis and chemokinesis. The translational strength of our in vitro BBB model was validated in vivo in mouse brains. We uncovered that CCR2 knock-down on tumor cells significantly reduces transmigration and can thus be harnessed as a potential therapeutic strategy to mitigate the early steps of the metastatic cascade at the brain. Furthermore, we expand upon the current BBB assay to recapitulate the complex tumor-stroma interactions with the incorporation of a high-grade glioma spheroid in the in vitro brain vasculature. Specifically, we explore the mechanisms of drug delivery, across the BBB and into the brain tumor, of layered nanoparticles that bind to endothelial receptors. With this novel platform and in vivo validation in glioma tumor-bearing mice, we demonstrate that transport occurs via transcytosis and is improved with LRP1-binding nanoparticles compared to control carriers, particularly across the vasculature near the glioma tumor. Keywords: cancer, extravasation, blood-brain barrier, microfluidics, organ-on-a-chip, glioma, nanoparticle
| Author | : Cynthia Hajal |
| Publisher | : |
| Total Pages | : 58 |
| Release | : 2018 |
| Genre | : |
| ISBN | : |
With up to 40% of cancer patients showing metastatic lesions to the brain and a 30% five-year survival rate post-diagnosis, secondary tumors to the brain are a leading cause of cancer-related deaths. Understanding the mechanisms of tumor cell extravasation at the brain is therefore crucial to the development of therapeutic agents targeting this step in cancer metastasis, and to the overall improvement of cancer survival rates . Investigating the interactions between tumor cells and brain stroma is of particular interest due to the site's unique microenvironment. In fact, the interface between brain and blood, known as the blood-brain barrier (BBB), is the tightest endothelial barrier in humans. The presence of tight junctions between brain endothelial cells, coupled with the spatial organization of pericytes and astrocytes around the vasculature, restrict the entry of most solutes and cells into the brain. Yet, the brain constitutes a common metastatic site to many primary cancers originating from the lung, breast and skin. This suggests that tumor cells must employ specific mechanisms to cross the blood-brain barrier. While in vitro models aimed at replicating the human blood-brain barrier exist, most are limited in their physiological relevance. In fact, the majority of these platforms rely on a monolayer of human brain endothelial cells in contact with pencytes, astrocytes and neurons. While this approach focuses on incorporating the relevant cell types of the brain microenvironment, it fails to accurately replicate the geometry of brain capillaries, the barrier tightness of the BBB, and the juxtacrine and paracrine signaling events occurring between brain endothelial cells and stromal cells during vasculogenesis. To integrate these features into a physiologically relevant blood-brain barrier model, we designed an in vitro microvascular network platform formed via vasculogenesis, using endothelial cells derived from human induced pluripotent stem cells, primary human brain pericytes, and primary human brain astrocytes. The vasculatures formed with brain pericytes and astrocytes exhibit decreased cross-section areas, increased endothelial cell-cell tight junction expression and basement membrane deposition, as well as reduced and more physiologically relevant values of vessel permeability, compared to the vasculatures formed with endothelial cells alone. The addition of pericytes and astrocytes in the vascular system was also coupled with increased extravasation efficiencies of different tumor cell subpopulations, despite the lower permeability values measured in this BBB model. Moreover, an increase in the extravasation potential of metastasized breast tumor cells collected from the brain was recorded with the addition of pericytes and astrocytes, with respect to the parental breast tumor cell line. These results were not observed in metastasized breast tumor cells collected from the lung, thus validating our BBB model and providing useful insight into the role of pericytes and astrocytes in extravasation. Our microfluidic platform certainly provides advantages over the current state-of-the-art in vitro blood-brain barrier models. While being more physiologically relevant than most in vitro platforms when it comes to geometry, barrier function and juxtacrine/paracrine signaling between the relevant cell types, our model provides a robust platform to understand tumor cell-brain stromal cell interactions during extravasation.
| Author | : Shay Soker |
| Publisher | : Humana Press |
| Total Pages | : 225 |
| Release | : 2017-10-20 |
| Genre | : Medical |
| ISBN | : 3319605119 |
Cancer cell biology research in general, and anti-cancer drug development specifically, still relies on standard cell culture techniques that place the cells in an unnatural environment. As a consequence, growing tumor cells in plastic dishes places a selective pressure that substantially alters their original molecular and phenotypic properties.The emerging field of regenerative medicine has developed bioengineered tissue platforms that can better mimic the structure and cellular heterogeneity of in vivo tissue, and are suitable for tumor bioengineering research. Microengineering technologies have resulted in advanced methods for creating and culturing 3-D human tissue. By encapsulating the respective cell type or combining several cell types to form tissues, these model organs can be viable for longer periods of time and are cultured to develop functional properties similar to native tissues. This approach recapitulates the dynamic role of cell–cell, cell–ECM, and mechanical interactions inside the tumor. Further incorporation of cells representative of the tumor stroma, such as endothelial cells (EC) and tumor fibroblasts, can mimic the in vivo tumor microenvironment. Collectively, bioengineered tumors create an important resource for the in vitro study of tumor growth in 3D including tumor biomechanics and the effects of anti-cancer drugs on 3D tumor tissue. These technologies have the potential to overcome current limitations to genetic and histological tumor classification and development of personalized therapies.
