scholarly journals Isolation and Characterization of Small Extracellular Vesicles from Porcine Blood Plasma, Cerebrospinal Fluid, and Seminal Plasma

Proteomes ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 17 ◽  
Author(s):  
Skalnikova ◽  
Bohuslavova ◽  
Turnovcova ◽  
Juhasova ◽  
Juhas ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Blood Plasma ◽  
Extracellular Vesicles ◽  
Seminal Plasma ◽  
Biomedical Research ◽  
Body Fluids ◽  
Large Animal ◽  
Large Animal Models ◽  
Isolation And Characterization ◽  
Proteomic Analyses

Extracellular vesicles (EVs) are a highly attractive subject of biomedical research as possible carriers of nucleic acid and protein biomarkers. EVs released to body fluids enable indirect access to inner organs by so-called “liquid biopsies”. Obtaining a high-quality EV sample with minimum contaminants is crucial for proteomic analyses using LC–MS/MS or other techniques. However, the EV content in various body fluids largely differs, which may hamper subsequent analyses. Here, we present a comparison of extracellular vesicle yields from blood plasma, cerebrospinal fluid, and seminal plasma using an experimental pig model. Pigs are widely used in biomedical research as large animal models with anatomy and physiology close to those of humans and enable studies (e.g., of the nervous system) that are unfeasible in humans. EVs were isolated from body fluids by differential centrifugation followed by ultracentrifugation. EVs were characterized according to protein yields and to the quality of the isolated vesicles (e.g., size distribution, morphology, positivity for exosome markers). In our experimental setting, substantial differences in EV amounts were identified among body fluids, with the seminal plasma being the richest EV source. The yields of pellet proteins from ultracentrifugation of 1 mL of porcine body fluids may help to estimate body fluid input volumes to obtain sufficient samples for subsequent proteomic analyses.

Livestock in biomedical research: history, current status and future prospective

2016 ◽  
Vol 28 (2) ◽  
pp. 112 ◽  
Author(s):  
Irina A. Polejaeva ◽  
Heloisa M. Rutigliano ◽  
Kevin D. Wells
Keyword(s):  
Genome Sequence ◽  
Biomedical Research ◽  
Assisted Reproductive Technologies ◽  
Sequence Data ◽  
Embryonic Stem ◽  
Reproductive Technologies ◽  
Farm Animals ◽  
Current Status ◽  
Large Animal ◽  
Large Animal Models

Livestock models have contributed significantly to biomedical and surgical advances. Their contribution is particularly prominent in the areas of physiology and assisted reproductive technologies, including understanding developmental processes and disorders, from ancient to modern times. Over the past 25 years, biomedical research that traditionally embraced a diverse species approach shifted to a small number of model species (e.g. mice and rats). The initial reasons for focusing the main efforts on the mouse were the availability of murine embryonic stem cells (ESCs) and genome sequence data. This powerful combination allowed for precise manipulation of the mouse genome (knockouts, knockins, transcriptional switches etc.) leading to ground-breaking discoveries on gene functions and regulation, and their role in health and disease. Despite the enormous contribution to biomedical research, mouse models have some major limitations. Their substantial differences compared with humans in body and organ size, lifespan and inbreeding result in pronounced metabolic, physiological and behavioural differences. Comparative studies of strategically chosen domestic species can complement mouse research and yield more rigorous findings. Because genome sequence and gene manipulation tools are now available for farm animals (cattle, pigs, sheep and goats), a larger number of livestock genetically engineered (GE) models will be accessible for biomedical research. This paper discusses the use of cattle, goats, sheep and pigs in biomedical research, provides an overview of transgenic technology in farm animals and highlights some of the beneficial characteristics of large animal models of human disease compared with the mouse. In addition, status and origin of current regulation of GE biomedical models is also reviewed.

Isolation and characterization of extracellular vesicles

2019 ◽  
Vol 1 (1) ◽  
pp. 18-26
Author(s):  
Fabia Fricke ◽  
Dominik Buschmann ◽  
Michael W. Pfaffl
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Body Fluids ◽  
Cell Culture Media ◽  
Advantages And Disadvantages ◽  
The Past ◽  
Isolation And Characterization ◽  
Basic Biology

Research into extracellular vesicles (EVs) gained significant traction in the past decade and EVs have been investigated in a wide variety of studies ranging from basic biology to diagnostic and therapeutic applications. Since EVs are secreted by most, if not all, eukaryotic and prokaryotic cells, they have been detected in body fluids as diverse as blood, urine and saliva as well as in cell culture media. In this chapter, we will provide an overview of EV isolation and characterization strategies and highlight their advantages and disadvantages.

Nucleotide pyrophosphatase and phosphodiesterase. I. Organ distribution and activities in body fluids.

1977 ◽  
Vol 23 (9) ◽  
pp. 1531-1537 ◽  
Author(s):  
H F Haugen ◽  
S Skrede
Keyword(s):  
Cerebrospinal Fluid ◽  
Seminal Plasma ◽  
Blood Cells ◽  
Body Fluids ◽  
Organ Distribution ◽  
Ph Optimum ◽  
Human Organs ◽  
Intestinal Lymph ◽  
Nucleotide Pyrophosphatase ◽  
Kidney Liver

Abstract We estimated nucleotide pyrophosphatase and phosphodiesterase I activities in human and rat organs and in body fluids from man and dog. The highest organ activities were found in epididymis, kidney, liver, and intestine. In body fluids, the activity was highest in seminal plasma, followed by intestinal lymph, serum, heart lymph, cerebrospinal fluid, milk, and urine. The ratio nucleotide pyrophosphatase/phosphodiesterase I and the urea resistance of phosphodiesterase I differed among human organs, body fluids, and blood cells. Different isoenzymes probably exist. The activities in serum share several properties with those in several organs--e.g. pH-optimum 9.6-9.8, dependency on Zn2+, and the effects of inhibitors. Phosphodiesterase I in erythrocytes, which has not been described previously, differs from enzyme from other sources by lower pH optimum (8.5), dependency on Mg2+, inhibition by Zn2+, and stimulation by dithiothreitol.

Characteristics and Unmet Clinical Needs Related to Hepatocellular Carcinoma

2017 ◽  
Vol 01 (02) ◽  
pp. 074-082
Author(s):  
Jordan Newson ◽  
Nickolas Kinachtchouk ◽  
Kyle Schachtschneider ◽  
Regina Schwind ◽  
Lawrence Schook
Keyword(s):  
Hepatocellular Carcinoma ◽  
Animal Models ◽  
Biomedical Research ◽  
Human Disease ◽  
Rabbit Model ◽  
In Vitro Study ◽  
Large Animal ◽  
Curative Therapy ◽  
Large Animal Models

AbstractAdvances in biomedical research require animal models that accurately recapitulate human disease. Without such models, progress against human diseases such as cancer is significantly hindered. Here, we present the current landscape on available and emerging hepatocellular carcinoma (HCC) animal models. HCC is the second leading cause of cancer death worldwide, with an annual death toll exceeding 745,000. Stunningly, only 15% of HCC patients are candidates for curative therapy, leading 85% of patients to seek palliative therapeutic options. The VX2 rabbit model is considered the most relevant and widely used HCC model; however, more reliable HCC models are critically needed. In general, animal models for biomedical research should (1) mimic the human disease on a molecular basis, (2) derive from a relevant cell line that lends itself to in vitro study, (3) be reliable and predictable, (4) manifest survival differences, (5) allow for accurate treatment assessment, (6) be readily imaged, and (7) occur in similar background settings as the human disease. Over the past decades, numerous small animal models have been utilized for HCC studies; however, the development of new large animal models as qualified alternatives to murine models represents a key technology to advance research into human clinical trials.

scholarly journals Use and Efficacy of Analgesic Agents in Sheep (Ovis aries) Used in Biomedical Research

2019 ◽  
Vol 58 (6) ◽  
pp. 755-766
Author(s):  
Mark W Stillman ◽  
Alexandra L Whittaker
Keyword(s):  
Biomedical Research ◽  
Local Anesthetics ◽  
Analgesic Effect ◽  
Nmda Antagonist ◽  
Ovis Aries ◽  
Large Animal ◽  
Channel Blockers ◽  
General Anesthetics ◽  
General Anesthetic ◽  
Large Animal Models

Sheep (Ovis aries) are widely used as large animal models in biomedical research. However, current literature on the use of analgesics in sheep generally focuses on an industry or farm level of use. This structured review evaluates use and efficacy of analgesics administered to sheep in a biomedical research setting. Electronic databases were searched with terms related to analgesia in research sheep. After application of exclusion criteria, 29 peer-reviewed publications were evaluated from 1995 to 2018. Drugs used for analgesia in sheep include opioids, α2 agonists, NSAID, local anesthetics, NMDA receptor antagonists, and calcium channel blockers. Opioid agonists have previously been considered short acting and of questionable efficacy in sheep, but newer modalities may provide effective analgesia. NSAID may exhibit an analgesic effect only when inflammatory pain is present and may not be beneficial for use in acute pain models. α2 agonists provide effective yet short-lived analgesia; however, side effects are of concern. Local anesthetics were previously widely used as stand-alone agents, as alternatives to the use of general anesthetics in sheep. These agents have since fallen out of favor as sole agents. Despite this, they provide a valuable analgesic effect when used as adjuncts to general anesthetic regimes. The NMDA antagonist ketamine provided good analgesia and is likely underutilized as an analgesic agent in sheep. Future controlled studies should further evaluate the analgesic properties of ketamine in sheep.

scholarly journals Compartmentalization and Antiviral Effect of Efavirenz Metabolites in Blood Plasma, Seminal Plasma, and Cerebrospinal Fluid

2012 ◽  
Vol 41 (2) ◽  
pp. 422-429 ◽  
Author(s):  
Lindsay B. Avery ◽  
Jennifer L. VanAusdall ◽  
Craig W. Hendrix ◽  
Namandjé N. Bumpus
Keyword(s):  
Cerebrospinal Fluid ◽  
Blood Plasma ◽  
Seminal Plasma ◽  
Antiviral Effect ◽  
Plasma Séminal ◽  
In Blood Plasma

scholarly journals Adipose-Derived Stromal/Stem Cells from Large Animal Models: from Basic to Applied Science

2020 ◽  
Author(s):  
Joanna Bukowska ◽  
Anna Zuzanna Szóstek-Mioduchowska ◽  
Marta Kopcewicz ◽  
Katarzyna Walendzik ◽  
Sylwia Machcińska ◽  
...  
Keyword(s):  
Stem Cells ◽  
Animal Models ◽  
Regenerative Medicine ◽  
Animal Health ◽  
Healing Process ◽  
Current Status ◽  
Wound Healing Process ◽  
Large Animal ◽  
Large Animal Models ◽  
Isolation And Characterization

Abstract Adipose-derived stem cells (ASCs) isolated from domestic animals fulfill the qualitative criteria of mesenchymal stem cells, including the capacity to differentiate along multiple lineage pathways and to self-renew, as well as immunomodulatory capacities. Recent findings on human diseases derived from studying large animal models, have provided evidence that administration of autologous or allogenic ASCs can improve the process of healing. In a narrow group of large animals used in bioresearch studies, pigs and horses have been shown to be the best suited models for study of the wound healing process, cardiovascular and musculoskeletal disorders. To this end, current literature demonstrates that ASC-based therapies bring considerable benefits to animal health in both spontaneously occurring and experimentally induced clinical cases. The purpose of this review is to provide an overview of the diversity, isolation, and characterization of ASCs from livestock. Particular attention has been paid to the functional characteristics of the cells that facilitate their therapeutic application in large animal models of human disease. In this regard, we describe outcomes of ASCs utilization in translational research with pig and horse models of disease. Furthermore, we evaluate the current status of ASC-based therapy in veterinary practice, particularly in the rapidly developing field of equine regenerative medicine. In conclusion, this review presents arguments that support the relevance of animal ASCs in the field of regenerative medicine and it provides insights into the future perspectives of ASC utilization in animal husbandry. Graphical abstract

scholarly journals Extracellular Vesicles in Blood: Sources, Effects and Applications

2021 ◽  
Vol 22 (15) ◽  
pp. 8163
Author(s):  
Ainhoa Alberro ◽  
Leire Iparraguirre ◽  
Adelaide Fernandes ◽  
David Otaegui
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Blood Cells ◽  
Circulatory System ◽  
Cell Types ◽  
Treatment Monitoring ◽  
Body Fluids ◽  
Comprehensive Overview ◽  
Isolation And Characterization ◽  
Almost All

Extracellular vesicles (EVs) are important players for intercellular communication. EVs are secreted by almost all cell types; they can transfer information between nearby or distant cells, and they are highly abundant in body fluids. In this review, we describe the general characteristics of EVs, as well as isolation and characterization approaches. Then, we focus on one of the most relevant sources of EVs: the blood. Indeed, apart from EVs secreted by blood cells, EVs of diverse origins travel in the bloodstream. We present the numerous types of EVs that have been found in circulation. Besides, the implications of blood-derived EVs in both physiological and pathological processes are summarized, highlighting their potential as biomarkers for the diagnosis, treatment monitoring, and prognosis of several diseases, and also as indicators of physiological modifications. Finally, the applications of EVs introduced in the circulatory system are discussed. We describe the use of EVs from distinct origins, naturally produced or engineered, autologous, allogeneic, or even from different species and the effects they have when introduced in circulation. Therefore, the present work provides a comprehensive overview of the components, effects, and applications of EVs in blood.

The size of extracellular vesicles secreted by different types of stem cells

2018 ◽  
pp. 38-42
Author(s):  
И.Б. Алчинова ◽  
М.В. Полякова ◽  
И.Н. Сабурина ◽  
М.Ю. Карганов
Keyword(s):  
Stem Cells ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Living Organism ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Study Results ◽  
Multipotent Mesenchymal Stem Cells ◽  
Rat Adipose Tissue ◽  
Almost All

Механизм терапевтического действия мультипотентных мезенхимных стволовых клеток (ММСК) на облученный организм в последнее время вызывает повышенный интерес исследователей. В качестве активного участника паракринного механизма реализации этого эффекта предлагают рассматривать внеклеточные везикулы, секретируемые практически всеми клетками живого организма. Цель работы: выделить и охарактеризовать внеклеточные везикулы, продуцируемые стволовыми клетками различной природы. Материалы и методы. Суспензии внеклеточных везикул, выделенных по модифицированному протоколу дифференциального центрифугирования из культуральных жидкостей от культур ММСК костного мозга человека 2-го пассажа и ММСК жировой ткани крысы 4-го пассажа, были проанализированы методом просвечивающей электронной микроскопии и методом анализа траекторий наночастиц. Результаты. Исследование показало наличие в обоих образцах микрочастиц размерами до и около 100 нм, однако процентное содержание частиц разных размеров в суспензии различалось для двух анализируемых типов клеток. Заключение. Полученные результаты могут свидетельствовать о специфике секреции, обусловленной клеточным типом. A mechanism of the therapeutic effect of multipotent mesenchymal stem cells (MMSC) on irradiated body has recently arisen much interest of researchers. Extracellular vesicles (EVs) secreted by almost all cells of a living organism were suggested to actively contribute to the paracrine mechanism of this effect. The aim of the study was isolation and characterization of extracellular vesicles produced by various types of stem cells. Materials and methods. Suspensions of EVs were isolated from culture media of passage 2 human bone marrow-derived MMSC and passage 4 rat adipose tissue-derived MMSC using a modified protocol of differential centrifugation and then studied using transmission electron microscopy and nanoparticle tracking analysis. Results. The study showed the presence of microparticles with a size of >100 nm in the examined samples. However, the percent content of particles with different sizes in the suspension was different in two analyzed types of cell culture. Conclusion. The study results might reflect a specificity of secretion determined by the cell type.

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