يتم تمويل ليون تاي من جامعة إلينوي شيكاغو بدء الأموال.

<ol>
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الكتاب ليس لديهم ما يكشف.

الأساليب المذكورة يمكن استخدامها لمعالجة العديد من الأسئلة البيولوجية الأساسية المحيطة بالتغطية الدماغية الوعائية 24 . على وجه التحديد، فإنها يمكن أن تحدد أي المستقبلات ومسارات الإشارات تلعب دورا في الأوعية الدموية، وتغطية السفينة في أنسجة السرطان، و?...

<html> الحاجز الدموي الدماغي (بب) من الشعيرات الدموية الدماغية هو أكبر واجهة من الدم إلى الدماغ الاتصال ويلعب دورا رئيسيا في التوازن من الجهاز العصبي المركزي (نس) 1 ، 2 . العمليات الحيوية في بب منع امتصاص الجزيئات غير المرغوب فيها من الدم، وإزالة النفايات من الجهاز العصبي المركزي، وتوفير المواد الغذائية الأساسية وجزيئات الإشارة إلى الجهاز العصبي المركزي، وتعديل نيوروينفلاماشيون 1 ، 2 ، 3 ، 4 ، 5 . تلف بب هو السائد خلال الشيخوخة والعديد من الاضطرابات العصبية بما في ذلك مرض الزهايمر (أد)، والتصلب المتعدد والسكتة الدماغية 1 ، 2 ، 3 ، 4 ، 5 ،أس = &quot;كريف&quot;&gt; 6. لذلك، قد تلعب خلل بب دورا رئيسيا في الاضطرابات العصبية، بما في ذلك كهدف علاجي. الحفاظ على تغطية السفينة مهم لوظائف التماثل من بب. ومع ذلك، في الجسم الحي وفي المختبر تضارب البيانات على ما إذا كانت العمليات التي تنطوي على الاضطرابات العصبية تسبب أعلى أو أقل تغطية بب 6 ، 7 ، 8 ، 9 ، 10 ، 11 ، 12 ، 13 ، وخاصة في م. لذلك، هناك مبرر قوي لتطوير نماذج في المختبر باستخدام أنواع الخلايا ذات الصلة لتقييم وفهم شامل لديناميات تغطية بب. وتتكون الشعيرات الدموية الدماغي من الخلايا النجمية، بيريسيتس والخلايا البطانية الدماغ (بيكس) <sأوب كلاس = &quot;كريف&quot;&gt; 3. تساهم جميع أنواع الخلايا في وظيفة بب من خلال الدعم الهيكلي وعن طريق إفراز جزيئات المستجيب مثل عوامل نمو الأوعية الدموية، السيتوكينات والكيموكينات التي تعمل بطريقة باراكرين و أوتوكرين مثل. ومع ذلك، فإن الخلايا المستجيب الرئيسية لل بب هي بيكس 3 . بشكل عام، وتقنيات زراعة الخلايا لتقييم وظيفة بب هي مقايسات نفاذية أجريت على الخلايا التي تزرع على إدراج فلتر، أو تقييم مستويات البروتينات الرئيسية بيك، سواء بعد إضافة الضغوطات 14 ، 15 ، 16 . على الرغم من أهمية، هذه المقايسات لا تركز على التغطية الدماغية الوعائية. هنا، يتم تفصيل أساليبنا السابقة 17 لتقييم تغطية بيك والهياكل الشبيهة بالشبكات باستخدام ثقافات واحدة من بيكس الإنسان الخالدة، والثقافات واحدة من بيك الأساسي الماوس، وثلاثة ثقافة إنسانيةنموذج (بيكس، أستروسيتس و بيريسيتس) من بب. وكان الهدف هو إظهار الأثر الضار لل OAβ، الذي يعتبر مساهما هاما في تقدم أد، على تغطية بيك. تأثير وقائي من عامل نمو البشرة (إغف) يسلط الضوء على إمكانات هذه التقنية كأداة الفحص العلاجي. هذه التقنية لديها العديد من التطبيقات واسعة للبحوث الأساسية والتطبيقية بما في ذلك: 1) تحديد دور مسارات محددة على الأوعية الدموية وتغطية السفينة، 2) تقييم آثار المرض والشيخوخة ذات الصلة العوامل على الأوعية الدموية وتغطية السفينة، و 3) تحديد الدوائية أهداف.

<table><tbody><tr><td>hCMEC/D3 cells</td><td>Milipore</td><td>SCC066</td><td></td></tr><tr><td>EBM-2 basal media</td><td>Lonza</td><td>CC-3156</td><td></td></tr><tr><td>Collagen Type 1</td><td>ThermoFisher</td><td>A1064401</td><td></td></tr><tr><td>HBSS, calcium, magnesium, no phenol red</td><td>ThermoFisher</td><td>14025092</td><td></td></tr><tr><td>HBSS, no calcium, no magnesium, no phenol red</td><td>ThermoFisher</td><td>14175095</td><td></td></tr><tr><td>Trypsin-EDTA (0.25%)</td><td>ThermoFisher</td><td>25200056</td><td></td></tr><tr><td>Final concentrations of the SingleQuot growth factor supplements for EBM2 media</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>5% FBS</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>10% Ascorbic acid</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>10% Gentamycin sulphate</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>25% Hydrocortisone</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>1/4 volume of the supplied growth factors: fibroblast growth factor, epidermal growth factor, insulin-like growth factor, vascular endothelial growth factor</td><td>Lonza</td><td>CC-4147</td><td></td></tr><tr><td>Puromycin hydrochloride</td><td>VWR</td><td>80503-312</td><td></td></tr><tr><td>MEM-HEPES</td><td>Thermo Scientific</td><td>12360-038</td><td></td></tr><tr><td>Papain cell dissociation system (papain and DNase1)</td><td>Worthington Biochemical</td><td>LK003150</td><td></td></tr><tr><td>Human pericytes</td><td>Sciencell</td><td>1200</td><td></td></tr><tr><td>Pericyte basal media</td><td>Sciencell</td><td>1201</td><td></td></tr><tr><td>Pericyte growth supplement</td><td>Sciencell</td><td>1252</td><td></td></tr><tr><td>Human Astrocytes</td><td>Sciencell</td><td>1800</td><td></td></tr><tr><td>Astrocyte media</td><td>Sciencell</td><td>1801</td><td></td></tr><tr><td>Astrocyte growth supplement</td><td>Sciencell</td><td>1852</td><td></td></tr><tr><td>Basement membrane (Matrigel Growth Factor Reduced)</td><td>Corning</td><td>356231</td><td></td></tr><tr><td>Angiogenesis m-plates (96-well)</td><td>ibidi</td><td>89646</td><td></td></tr><tr><td>Human Epidermal growth factor</td><td>Shenendoah Biotechnology</td><td>100-26</td><td></td></tr><tr><td>CellTracker green</td><td>ThermoFisher</td><td>C7025</td><td></td></tr><tr><td>CellTracker orange</td><td>ThermoFisher</td><td>C34551</td><td></td></tr><tr><td>CellTracker blue</td><td>ThermoFisher</td><td>C2110</td><td></td></tr><tr><td>Poly-l-lysine</td><td>Sciencell</td><td>0403</td><td></td></tr><tr><td>10% Neutral Buffered Formalin</td><td>Sigma-Aldrich</td><td>HT5012-60ML</td><td></td></tr><tr><td>C57BL mice</td><td>Jackson Laboratory</td><td>na</td><td></td></tr><tr><td>PCR tube strips</td><td>GeneMate</td><td>T-3014-2</td><td></td></tr><tr><td>Zeiss stereo discover v.8 dissecting microscope</td><td>Zeiss</td><td>na</td><td></td></tr></tbody></table>

in vitro assays to assess blood-brain barrier mesh-like vessel formation and disruption

تلعب حاجز الدم في الدماغ (بب) دورا محوريا في توازن الجهاز العصبي المركزي (نس). يتم الحفاظ على حيوي بب بواسطة الخلايا النجمية، بيريسيتس والخلايا البطانية في الدماغ (بيكس). هنا، نحن بالتفصيل أساليب لتقييم تغطية بب باستخدام الثقافات واحدة من بيكس الإنسان الخالد، والثقافات واحدة من بيك الأساسي الماوس، ونموذج الثقافة الثلاثي أنسنة (بيكس، النجمية و بيريسيتس) من بب. لتسليط الضوء على تطبيق المقايسات لحالات المرض، ونحن تصف تأثير أميلويد أوليغوميريك β (oAβ)، الذي هو مساهم مهم في مرض الزهايمر (أد) التقدم، على تغطية بب. وعلاوة على ذلك، فإننا نستخدم عامل نمو البشرة (إغف) لإلقاء الضوء على إمكانية فحص المخدرات من التقنيات. نتائجنا تبين أن واحدة وثلاثية بيكس مثقف تشكل هياكل تشبه شبكة في ظل الظروف القاعدية، وأن oAβ يعطل هذه التشكيل خلية تشكيل ويتحول الهياكل شبكة بريفورمد،ولكن إغف يمنع هذا الاضطراب. وبالتالي، فإن التقنيات الموصوفة مهمة لتشريح العمليات الأساسية والمرض ذات الصلة التي تعدل تغطية بب.

في الثقافات واحدة، على حد سواء خلايا هميك / D3 ( الشكل 3A ) و بيس الأساسي الماوس ( الشكل 3B ) شكل هياكل تشبه شبكة في جميع أنحاء البئر. وتتميز الهياكل بشبكة من العقد المترابطة ( الشكل 3 ). في جميع النماذج وصفها ( ...

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In Vitro Assays to Assess Blood-brain Barrier Mesh-like Vessel Formation and Disruption

الحفاظ على تغطية حاجز الدم في الدماغ هو المفتاح لتوازن الجهاز العصبي المركزي. يصف هذا البروتوكول في تقنيات المختبر لترسيم العمليات الأساسية والمرضية التي تعدل تغطية حاجز الدم في الدماغ.

في مقايسات المختبر لتقييم الحاجز الدموي الدماغي تشبه تشكيل سفينة وتشوه

Blood-brain barrier (BBB) coverage plays a central role in the homeostasis of the central nervous system (CNS). The BBB is dynamically maintained by ...

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In Vitro Assays to Assess Blood-brain Barrier Mesh-like Vessel Formation and Disruption

8.5K Views.  University of Illinois at Chicago.  الحفاظ على تغطية حاجز الدم في الدماغ هو المفتاح لتوازن الجهاز العصبي المركزي. يصف هذا البروتوكول في تقنيات المختبر لترسيم العمليات الأساسية والمرضية التي تعدل تغطية حاجز الدم في الدماغ. 

Video: In Vitro Assays to Assess Blood-brain Barrier Mesh-like Vessel Formation and Disruption

Predicting In Vivo Payloads Delivery using a Blood-brain Tumor-barrier in a Dish

Highly selective by nature, the blood-brain barrier (BBB) is essential for the brain homeostasis in physiological conditions. However, in the context of brain tumors, the molecular selectivity of BBB also shields the neoplastic cells by blocking the delivery of peripherally administered chemotherapies. The development of novel drugs (including nanoparticles) targeting malignant brain tumors ideally requires the use of preclinical animal models to study the drug&#8217;s transcytosis and antitumor efficacy. In order to comply with the 3R principle (refine, reduce, and replace) to reduce the number of laboratory animals in experimental setup and perform the high-throughput screening of a large library of antitumor agents, we developed a reproducible in vitro human and murine mimic of the blood-brain tumor-barrier (BBTB) using three-layered cultures of endothelial cells, astrocytes, and patient-derived glioblastoma spheres. For higher scalability and reproducibility, commercial cell lines or immortalized cells have been used in tailored conditions to allow the formation of a barrier resembling the actual BBB. Here we describe a protocol to obtain a BBTB mimic by culturing endothelial cells in contact with astrocytes at specific cell densities on inserts. This BBTB mimic can be used, for instance, for the quantification and confocal imaging of the nanoparticle passage through the endothelial and astrocytic barriers, in addition to the evaluation of the tumor cell targeting within the same assay. Moreover, we show that the obtained data can be used to predict the behavior of nanoparticles in preclinical animal models. In a broader perspective, this in vitro model could be adapted to other neurodegenerative diseases for the determination of the passage of new therapeutic molecules through the BBB and/or be supplemented with brain organoids to directly evaluate the efficacy of drugs.

Drug targeting to central nervous system tumors is a major challenge. Here we describe a protocol to produce an in vitro mimic of the blood-brain tumor-barrier using murine and/or human cells and discuss their relevance for the predictability of central nervous system tumor targeting in vivo.

التنبؤ في فيفو حمولات التسليم استخدام ورم-حاجز دم في الدماغ في صحن

Highly selective by nature, the blood-brain barrier (BBB) is essential for the brain homeostasis in physiological conditions. However, in the context of ...

Cancer Research

Improved Method for the Preparation of a Human Cell-based, Contact Model of the Blood-Brain Barrier

The blood-brain barrier (BBB) comprises impermeable but adaptable brain capillaries which tightly control the brain environment. Failure of the BBB has been implied in the etiology of many brain pathologies, creating a need for development of human in vitro BBB models to assist in clinically-relevant research. Among the numerous BBB models thus far described, a static (without flow), contact BBB model, where astrocytes and brain endothelial cells (BECs) are cocultured on the opposite sides of a porous membrane, emerged as a simplified yet authentic system to simulate the BBB with high throughput screening capacity. Nevertheless the generation of such model presents few technical challenges. Here, we describe a protocol for preparation of a contact human BBB model utilizing a novel combination of primary human BECs and immortalized human astrocytes. Specifically, we detail an innovative method for cell-seeding on inverted inserts as well as specify insert staining techniques and exemplify how we use our model for BBB-related research.

Establishment of human models of the blood-brain barrier (BBB) can benefit research into brain conditions associated with BBB failure. We describe here an improved technique for preparation of a contact BBB model, which permits coculturing of human astrocytes and brain endothelial cells on the opposite sides of a porous membrane.

تحسين طريقة لإعداد القائم على الخليوي، الاتصال نموذج الإنسان من حاجز الدم في الدماغ

The blood-brain barrier (BBB) comprises impermeable but adaptable brain capillaries which tightly control the brain environment. Failure of the BBB has ...

Bioengineering

Setting-up an In Vitro Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated Transport

The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue. Our aim was to develop and characterize a highly reproducible rat syngeneic in vitro model of the BBB using co-cultures of primary rat brain endothelial cells (RBEC) and astrocytes to study receptors involved in transcytosis across the endothelial cell monolayer. Astrocytes were isolated by mechanical dissection following trypsin digestion and were frozen for later co-culture. RBEC were isolated from 5-week-old rat cortices. The brains were cleaned of meninges and white matter, and mechanically dissociated following enzymatic digestion. Thereafter, the tissue homogenate was centrifuged in bovine serum albumin to separate vessel fragments from nervous tissue. The vessel fragments underwent a second enzymatic digestion to free endothelial cells from their extracellular matrix. The remaining contaminating cells such as pericytes were further eliminated by plating the microvessel fragments in puromycin-containing medium. They were then passaged onto filters for co-culture with astrocytes grown on the bottom of the wells. RBEC expressed high levels of tight junction (TJ) proteins such as occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. The transendothelial electrical resistance (TEER) of brain endothelial monolayers, indicating the tightness of TJs reached 300 ohm&#183;cm2 on average. The endothelial permeability coefficients (Pe) for lucifer yellow (LY) was highly reproducible with an average of 0.26 &#177;&#160;0.11 x 10-3 cm/min. Brain endothelial cells organized in monolayers expressed the efflux transporter P-glycoprotein (P-gp), showed a polarized transport of rhodamine 123, a ligand for P-gp, and showed specific transport of transferrin-Cy3 and DiILDL across the endothelial cell monolayer. In conclusion, we provide a protocol for setting up an in vitro BBB model that is highly reproducible due to the quality assurance methods, and that is suitable for research on BBB transporters and receptors.

Setting-up an In Vitro Model of Rat Blood-brain Barrier BBB: A Focus on BBB Impermeability and Receptor-mediated Transport

The aim of the present study was to validate the reproducibility of an in vitro BBB model involving a rat syngeneic co-culture of endothelial cells and astrocytes. The endothelial cell monolayer presented high TEER and low LY permeability. Expression of specific TJ proteins, functional responses to inflammation and functionality of transporters and receptors were assessed.

وضع ما يصل إلى<em&gt; في المختبر</em&gt; نموذج من الفئران حاجز الدم في الدماغ (BBB): التركيز على BBB عدم النفاذ والنقل مستقبلات بوساطة

The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue. Our aim was to develop and ...

An In Vitro Model of the Blood-brain Barrier Using Impedance Spectroscopy: A Focus on T Cell-endothelial Cell Interaction

Breakdown of the blood-brain barrier (BBB) is a critical step in the development of autoimmune diseases such as multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). This process is characterized by the transmigration of activated T cells across brain endothelial cells (ECs), the main constituents of the BBB. However, the consequences on brain EC function upon interaction with such T cells are largely unknown. Here we describe an assay that allows for the evaluation of primary mouse brain microvascular EC (MBMEC) function and barrier integrity during the interaction with T cells over time. The assay makes use of impedance cell spectroscopy, a powerful tool for studying EC monolayer integrity and permeability, by measuring changes in transendothelial electrical resistance (TEER) and cell layer capacitance (Ccl). In direct contact with ECs, stimulated but not na&#239;ve T cells are capable of inducing EC monolayer dysfunction, as visualized by a decrease in TEER and an increase in Ccl. The assay records changes in EC monolayer integrity in a continuous and automated fashion. It is sensitive enough to distinguish between different strengths of stimuli and levels of T cell activation and it enables the investigation of the consequences of a targeted modulation of T cell-EC interaction using a wide range of substances such as antibodies, pharmacological reagents and cytokines. The technique can also be used as a quality control for EC integrity in in vitro T-cell transmigration assays. These applications make it a versatile tool for studying BBB properties under physiological and pathophysiological conditions.

An In Vitro Model of the Blood-brain Barrier Using Impedance Spectroscopy: A Focus on T Cell-endothelial Cell Interaction

Here, we describe an in vitro murine model of the blood-brain barrier that makes use of impedance cell spectroscopy, with a focus on the consequences on endothelial cell integrity and permeability upon interaction with activated T cells.

ل<em&gt; في المختبر</em&gt; نموذج من حاجز الدم في الدماغ عن طريق التحليل الطيفي المعاوقة: التركيز على التفاعل خلية تي خلية البطانية

Breakdown of the blood-brain barrier (BBB) is a critical step in the development of autoimmune diseases such as multiple sclerosis (MS) and its animal ...

Immunology and Infection

Analysis of Lymphocyte Extravasation Using an In Vitro Model of the Human Blood-brain Barrier

Lymphocyte extravasation into the central nervous system (CNS) is critical for immune surveillance. Disease-related alterations of lymphocyte extravasation might result in pathophysiological changes in the CNS. Thus, investigation of lymphocyte migration into the CNS is important to understand inflammatory CNS diseases and to develop new therapy approaches. Here we present an in vitro model of the human blood-brain barrier to study lymphocyte extravasation. Human brain microvascular endothelial cells (HBMEC) are confluently grown on a porous polyethylene terephthalate transwell insert to mimic the endothelium of the blood-brain barrier. Barrier function is validated by zonula occludens immunohistochemistry, transendothelial electrical resistance (TEER) measurements as well as analysis of evans blue permeation. This model allows investigation of the diapedesis of rare lymphocyte subsets such as CD56brightCD16dim/- NK cells. Furthermore, the effects of other cells, cytokines and chemokines, disease-related alterations, and distinct treatment regimens on the migratory capacity of lymphocytes can be studied. Finally, the impact of inflammatory stimuli as well as different treatment regimens on the endothelial barrier can be analyzed.

Analysis of Lymphocyte Extravasation Using an In Vitro Model of the Human Blood-brain Barrier

Here, we describe a human blood-brain barrier model enabling to investigate lymphocyte transmigration into the central nervous system in vitro.

تحليل الخلايا الليمفاوية التسرب باستخدام<em&gt; في المختبر</em&gt; نموذج من الدم في الدماغ الإنسان الحاجز

Lymphocyte extravasation into the central nervous system (CNS) is critical for immune surveillance. Disease-related alterations of lymphocyte ...

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains brain homeostasis. The principal sites of the barrier are endothelial cells of the brain capillaries whose barrier function results from tight intercellular junctions and efflux transporters expressed on the plasma membrane. This function is regulated by pericytes and astrocytes that together form the neurovascular unit (NVU). Several neurological diseases such as stroke, Alzheimer&#39;s disease (AD), brain tumors are associated with an impaired BBB function. Assessment of the BBB permeability is therefore crucial in evaluating the severity of the neurological disease and the success of the treatment strategies employed.
We present here a simple yet robust permeability assay that have been successfully applied to several mouse models both, genetic and experimental. The method is highly quantitative and objective in comparison to the tracer fluorescence analysis by microscopy that is commonly applied. In this method, mice are injected intraperitoneally with a mix of aqueous inert fluorescent tracers followed by anesthetizing the mice. Cardiac perfusion of the animals is performed prior to harvesting brain, kidneys or other organs. Organs are homogenized and centrifuged followed by fluorescence measurement from the supernatant. Blood drawn from the cardiac puncture just before perfusion serves for normalization purpose to the vascular compartment. The tissue fluorescence is normalized to the wet weight and serum fluorescence to obtain a quantitative tracer permeability index. For additional confirmation, the contralateral hemi-brain preserved for immunohistochemistry can be utilized for tracer fluorescence visualization purposes.

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Here we present a mouse brain vascular permeability assay using intraperitoneal injection of fluorescent tracers followed by perfusion that is applicable to animal models of blood-brain barrier dysfunction. One hemi-brain is used for assessing permeability quantitatively and the other for tracer visualization/immunostaining. The procedure takes 5 - 6 h for 10 mice.

في فيفو حاجز الدم في الدماغ نفاذية مقايسة في الفئران باستخدام المسمى فلوريسسينتلي تتبع

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains ...

Neuroscience

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 &#177; 80 &#937; cm2, and paracellular permeability (Papp) for Lucifer Yellow of 0.90 10-6 &#177; 0.13 10-6 cm sec-1 (mean &#177; SEM, n=55). Further evaluation of this pBEC phenotype showed good expression of the tight junctional proteins claudin 5, ZO-1, occludin and adherens junction protein p120 catenin. The model presented can be used for a range of studies of the BBB in health and disease and, with the highly restrictive paracellular permeability, this model is suitable for studies of transport and intracellular trafficking.

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

تحسين أسلوب لإنشاء في المختبر حاجز الدم – المخ نموذج استناداً إلى خلايا المخ الخنزيري بطانية

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

نموذج حاجز الدم في الدماغ 3D مشتق من الخلايا الجذعية البشرية متعددة القدرات

Reconstruction of the Blood-Brain Barrier In Vitro to Model and Therapeutically Target Neurological Disease

The blood-brain barrier (BBB) is a key physiological component of the central nervous system (CNS), maintaining nutrients, clearing waste, and protecting the brain from pathogens. The inherent barrier properties of the BBB pose a challenge for therapeutic drug delivery into the CNS to treat neurological diseases. Impaired BBB function has been related to neurological disease. Cerebral amyloid angiopathy (CAA), the deposition of amyloid in the cerebral vasculature leading to a compromised BBB, is a co-morbidity in most cases of Alzheimer&#39;s disease (AD), suggesting that BBB dysfunction or breakdown may be involved in neurodegeneration. Due to limited access to human BBB tissue, the mechanisms that contribute to proper BBB function and BBB degeneration remain unknown. To address these limitations, we have developed a human pluripotent stem cell-derived BBB (iBBB) by incorporating endothelial cells, pericytes, and astrocytes in a 3D matrix. The iBBB self-assembles to recapitulate the anatomy and cellular interactions present in the BBB. Seeding iBBBs with amyloid captures key aspects of CAA. Additionally, the iBBB offers a flexible platform to modulate genetic and environmental factors implicated in cerebrovascular disease and neurodegeneration, to investigate how genetics and lifestyle affect disease risk. Finally, the iBBB can be used for drug screening and medicinal chemistry studies to optimize therapeutic&#160;delivery to the CNS. In this protocol, we describe the differentiation of the three types of cells (endothelial cells, pericytes, and astrocytes) arising from human pluripotent stem cells, how to assemble the differentiated cells into the iBBB, and how to model CAA in vitro using exogenous amyloid. This model overcomes the challenge of studying live human brain tissue with a system that has both biological fidelity and experimental flexibility, and enables the interrogation of the human BBB and its role in neurodegeneration.

تسليط الضوء على المؤلف: نهج شخصي للتحقيق في مرض الزهايمر باستخدام نموذج حاجز الدم في الدماغ في المختبر 

The blood-brain barrier (BBB) has a crucial role in sustaining a stable and healthy brain environment. BBB dysfunction is associated with many neurological diseases. We have developed a 3D, stem-cell-derived model of the BBB to investigate cerebrovascular pathology, BBB integrity, and how the BBB is altered by genetics and disease.

إعادة بناء الحاجز الدموي الدماغي في المختبر لنمذجة الأمراض العصبية واستهدافها علاجيا

The blood-brain barrier (BBB) is a key physiological component of the central nervous system (CNS), maintaining nutrients, clearing waste, and protecting ...

في مقايسات المختبر لتقييم الحاجز الدموي الدماغي تشبه تشكيل سفينة وتشوه

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التنبؤ في فيفو حمولات التسليم استخدام ورم-حاجز دم في الدماغ في صحن

تحسين طريقة لإعداد القائم على الخليوي، الاتصال نموذج الإنسان من حاجز الدم في الدماغ

وضع ما يصل إلى

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تحليل الخلايا الليمفاوية التسرب باستخدام

في فيفو حاجز الدم في الدماغ نفاذية مقايسة في الفئران باستخدام المسمى فلوريسسينتلي تتبع

تحسين أسلوب لإنشاء في المختبر حاجز الدم – المخ نموذج استناداً إلى خلايا المخ الخنزيري بطانية

إعادة بناء الحاجز الدموي الدماغي في المختبر لنمذجة الأمراض العصبية واستهدافها علاجيا

إعادة بناء الحاجز الدموي الدماغي في المختبر لنمذجة الأمراض العصبية واستهدافها علاجيا

إعادة بناء الحاجز الدموي الدماغي في المختبر لنمذجة الأمراض العصبية واستهدافها علاجيا