الكتاب هم من موظفي شركة محدودة Miltenyi بيوتيك ، ألمانيا.

على العمل تحت ظروف معقمة ، فمن المستحسن لتنفيذ كافة الخطوات في تدفق الصفحي هود. 1. المواد <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> HEPES العازلة : 10 مم HEPES - هيدروكسيد الصوديوم الهيدروجيني 7.4 ، 150 مم كلوريد الصوديوم ، 5 ملي بوكل ، 1 ملم MgCl 2 و 1.8 ملي CaCl 2) </li><li style=";text-align:right;direction:rtl"> كولاجيناز D الحل : كولاجيناز D : 100 ملغ / مل (كولاجيناز D&gt; 0.15 U / ملغ) ، في HEPES العازلة </li><li style=";text-align:right;direction:rtl"> الدناز أنا الحل : 20،000 U / مل أنا الدناز </li><li style=";text-align:right;direction:rtl"> PBS العازلة عند الرقم الهيدروجيني 7.2 </li><li style=";text-align:right;direction:rtl"> PEB العازلة : 1 ألبوم جيش صرب البوسنة في 20 جزءا الحل أجزاء autoMACS الحل الشطف </li><li style=";text-align:right;direction:rtl"> نسيج : سرطان الرئة المستمدة من 07/06 الاسبوع القديمة الماوس الكبار BALB / C الإناث ، وخالية من الأجهزة المجاورة. </li><li style=";text-align:right;direction:rtl"> اختياري : CD11c MicroBeads ، MACS أعمدة MS ، MACS فاصل لعزل الخلايا الجذعية من الايقاف فأر وحيد الخلية الرئة </li></ol> 2. عدم الربط بين أنسجة الرئة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> شطف الأنسجة في طبق بتري تحتوي على برنامج تلفزيوني لإزالة كريات الدم الحمراء. </li><li style=";text-align:right;direction:rtl"> نقل كحد أقصى من 450 ملغ إلى أنسجة الرئة أنبوب C gentleMACS تحتوي على 4.9 مل HEPES العازلة. </li><li style=";text-align:right;direction:rtl"> إضافة 100 ميكرولتر حل D كولاجيناز إلى تركيز النهائي من 2 دال كولاجيناز ملغ / مل </li><li style=";text-align:right;direction:rtl"> أضف 10 ميكرولتر الحل الأول الدناز لمدة تصل إلى 150 ملغ الأنسجة (تركيز النهائي من 40 يو / مل الدناز I) أو 20 ميكرولتر الدناز I 150-450 ملغ لأنسجة الرئة (تركيز النهائي من 80 الدناز I U / مل). </li><li style=";text-align:right;direction:rtl"> إغلاق الأنبوب بإحكام C ونعلق وضعها على Dissociator gentleMACS. ثم تشغيل برنامج &quot;m_lung_01&quot;. </li><li style=";text-align:right;direction:rtl"> احتضان لمدة 30 دقيقة عند 37 درجة مئوية مع تناوب الآلي أو يدويا تحول كل 5 دقائق. </li><li style=";text-align:right;direction:rtl"> عودة إلى عينة Dissociator gentleMACS وتشغيل برنامج &quot;m_lung_02&quot;. </li></ol> 3. تصفية <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يعد أنبوب 50 مل لجمع الخلايا التي تمت تصفيتها من خلال استبدال الغطاء بمصفاة 70 ميكرومتر شبكة خلية. </li><li style=";text-align:right;direction:rtl"> مرة واحدة في برنامج gentleMACS الثانية بعد ، اعتمادا على توزيع المواد العينة في الأنبوب ، قد الطرد المركزي في أنبوب لفترة قصيرة لجمع مادة العينة في الجزء السفلي من الأنبوب. </li><li style=";text-align:right;direction:rtl"> إزالة الخلايا من خلال الغطاء الحاجز مختومة من الأنبوب C باستخدام ماصة مناسبة 1000 غيض ميكرولتر وتطبيقها على مصفاة الخلية. </li><li style=";text-align:right;direction:rtl"> غسل مصفاة الخلية مع 5 مل HEPES العازلة في RT. </li><li style=";text-align:right;direction:rtl"> الطرد المركزي الخلايا لبيليه في أنبوب مل 50 في 300xg لمدة 10 دقيقة. </li><li style=";text-align:right;direction:rtl"> نضح في وطاف resuspend الكرية في حجم الخلية التي تريدها ، PEB العازلة. </li></ol> الجزء 4 : النتائج الممثل : <img src="/files/ftp_upload/1266/1266fig1.jpg" alt="الشكل 1" /> الشكل 1. أسفرت الرئة تفارق مع Dissociator ™ gentleMACS في خلايا قابلة للحياة 92 ٪. وكانت ملطخة fluorescently الخلايا الميتة مع يوديد propidium (PI) (الحق مؤامرة نقطة ؛ اليسار مؤامرة نقطة : لا تلطيخ PI). <img src="/files/ftp_upload/1266/1266fig2.jpg" alt="الشكل 2" /> الشكل 2. تفكك أنسجة الرئة باستخدام الماوس نتائج Dissociator gentleMACS في ارتفاع نسبة الكريات البيض قابلة للحياة والخلايا البطانية. وكانت ملطخة المستمدة حيدة الخلية مع تعليق PE - CD45 و CD146 FITC أو CD31 FITC وتحليلها من قبل التدفق الخلوي. <img src="/files/ftp_upload/1266/1266figure2c.png" alt="الشكل 2C" /> وكان الرقم 3 الملون. حيدة الخلية تعليق المستمدة من أنسجة الرئة الفأر مع CD11c - FITC ومكافحة mPDCA APC - 1 - لكشف الماوس CD11c منخفضة M - PDCA - 1 + بلازماوية البلدان النامية وكذلك ارتفاع CD11c الخلايا. <img src="/files/ftp_upload/1266/1266fig4.jpg" alt="الشكل 4" /> الشكل 4. إثراء باستخدام الخلايا الجذعية MicroBeads CD11c ، وهو فاصل MiniMACS واثنين من أعمدة MS.

<ol>
	<li>Swanson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flt3-Ligand,+IL-4,+GM-CSF,+and+Adherence-Mediated+Isolation+of+Murine+Lung+Dendritic+Cells:+Assessment+of+Isolation+Technique+on+Phenotype+and+Function.">Flt3-Ligand, IL-4, GM-CSF, and Adherence-Mediated Isolation of Murine Lung Dendritic Cells: Assessment of Isolation Technique on Phenotype and Function.</a> J. Immunol. 173, 4875-4881 (2004).</li><li>Vermaelen, K., Pauwels, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+and+simple+discrimination+of+mouse+pulmonary+dendritic+cell+and+macrophage+populations+by+flow+cytometry:+Methodology+and+new+insights.">Accurate and simple discrimination of mouse pulmonary dendritic cell and macrophage populations by flow cytometry: Methodology and new insights.</a> Cytometry Part A. 61A, 170-177 (2004).</li></ol>

The authors are employees of Miltenyi Biotec GmbH who make the instrument used in this article.

في هذا الفيديو ، نقدم طريقة جديدة للتفكك أنسجة الرئة الماوس. نظهر أن الجمع بين المعالجة الميكانيكية والأنزيمية للأنسجة الرئة حققت نسبة عالية من الكريات البيض قابلة للحياة والخلايا البطانية. على وجه التحديد ، وقد تحقق تفكك الميكانيكية للأنسجة التي Dissociator gentleMACS. جيم أ...

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<tr>
<td>C Tubes</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-237</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx">http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx</a></td>
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<td>gentleMACS&#x2122; Starting Kit</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-235</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx">http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx</a></td>
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<td>MACSmix&#x2122; Tube Rotator</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-090-753</td>
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<td>Miltenyi Biotec</td>
<td>Order no. 130-091-222</td>
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<td>Miltenyi Biotec</td>
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<td>Miltenyi Biotec</td>
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<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-091-963</td>
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<td>Miltenyi Biotec</td>
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<td>Miltenyi Biotec</td>
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<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-233</td>
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<td>Art 1000 Reach Pipet Tips</td>
<td>&nbsp;</td>
<td>Molecular BioProducts</td>
<td>Order no. 2079</td>
<td><a href="http://www.mbpinc.com/ASP/products.aspx?query_TipID=61">http://www.mbpinc.com/ASP/products.aspx?query_TipID=61</a> <a href="http://www.mbpinc.com/html/products/art/display.html">http://www.mbpinc.com/html/products/art/display.html</a>
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standardized preparation of single-cell suspensions from mouse lung tissue using the gentlemacs dissociator

إعداد وحيدة الخلية تعليق من الأنسجة شرطا مسبقا هاما لكثير من التجارب في مجال البحوث الخلوية. عملية الفصل بين أجهزة كله يتطلب المعلمات محددة من أجل الحصول على عدد كبير من خلايا قابلة للحياة بطريقة استنساخه. وDissociator gentleMACS يحسن هذه المهمة مع بروتوكول بسيطة وعملية. الأداة مبرمجة مسبقا يحتوي على الإعدادات التي هي الأمثل للكفاءة ولكن لطيف تفارق من مجموعة متنوعة من أنواع الأنسجة ، بما في ذلك الرئتين الماوس. في هذا المنشور ويتجلى استخدام Dissociator gentleMACS على أنسجة الرئة المستمدة من الفئران.

Watch this Scientific Journal Video about Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator at JoVE.com

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

الفصل بين أنواع خلايا من نسيج معين يتطلب بارامترات محددة للتحريض الأنسجة للحصول على كميات كبيرة من الخلايا ، قابلة للحياة زروع. وgentleMACS Miltenyi dissociator يحسن هذه المهمة مع بروتوكول بسيطة وعملية. هذا المنشور هو شرح استخدام هذا الجهاز عن نسيج الرئة.

إعداد موحدة وحيدة الخلية معلقات من أنسجة الرئة باستخدام الماوس Dissociator gentleMACS

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

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34.3K Views.  Miltenyi Biotec,GmbH.  中文 日本語 

Video: Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual dissociations the gentleMACS Dissociator allows one to dissociate tissue very efficiently under controlled and reproducible conditions.  The gentleMACS Dissociator can optimally dissociate mouse spleen, combining timesaving and standardization with user-safety.

This video describes how to dissociate mouse spleens using the gentleMACS Dissociator, an automated bench-top device that can mechanically disrupt tissues using special tubes to produce viable cell suspensions. Following dissociation, spleens are filtered, centrifuged, and resuspended for further applications.

This video describes a simple, time-saving technique for automated tissue dissociation using the gentleMACS Dissociator to prepare single-cell suspensions of mouse splenocytes.

إعداد وحيدة الخلية معلقات من الطحال ماوس مع Dissociator gentleMACS

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual ...

Generation of Single-Cell Suspensions from Mouse Neural Tissue

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a repertoire of cell surface molecules, or molecular determinants, through which it can be identified and characterized. Currently, robust cell identification and separation technologies require single-cell preparations to be generated while simultaneously limiting cell death and destruction of characteristic surface protein. The gentleMACS Dissociator, when used in combination with trypsin or papain-based dissociation kits, can effectively and gently dissociate brain tissue while preserving antigen epitopes and limiting cell loss. Standardized preparation of single-cell suspensions is achieved using C Tubes and optimized, preset gentleMACS Programs. Once generated, single-cell suspensions can be treated with monoclonal conjugates like Anti-Prominin-1 MicroBeads, which identify neural progenitors, or purified further using Myelin Removal Beads.

Dissociating cells from specific tissue types requires specific parameters for tissue aggitation to obtain a high volume of viable, culturable cells. The Miltenyi gentleMACS Dissociator optimizes this task with a simple, practical protocol. In this publication the use of this apparatus on nerual tissue is explained.

جيل وحيدة الخلية معلقات من الأنسجة العصبية ماوس

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a ...

Single-cell Suction Recordings from Mouse Cone Photoreceptors

Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment are open and allow cations to flow steadily inwards across the membrane, depolarizing the cell. Light exposure triggers the closure of the CNG channels, blocks the inward cation current flow, and thus results in cell hyperpolarization. Based on the polarity of photoreceptors, a suction recording method was developed in 1970s that, unlike the classic patch-clamp technique, does not require penetrating the plasma membrane 1. Drawing the outer segment into a tightly-fitting glass pipette filled with extracellular solution allows recording the current changes in individual cells upon test-flash exposure. However, this well-established "outer-segment-in (OS-in)" suction recording is not suitable for mouse cone recordings, because of the low percentage of cones in the mouse retina (3%) and the difficulties in identifying the cone outer segments. Recently, an inner-segment-in (IS-in) recording configuration was developed to draw the inner segment/nuclear region of the photoreceptor into the recording pipette 2,3. In this video, we will show how to record from individual mouse cone photoresponses using single-cell suction electrode.

We will show how to record flash responses from single mouse cones using a suction electrode.

تسجيلات شفط وحيدة الخلية المستقبلة من المخروط ماوس

Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also produce a source of patient-matched cells for regenerative therapies. iPSCs may be generated using multiple protocols and derived from multiple cell sources. Once generated, iPSCs are tested using a variety of assays including immunostaining for pluripotency markers, generation of three germ layers in embryoid bodies and teratomas, comparisons of gene expression with embryonic stem cells (ESCs) and production of chimeric mice with or without germline contribution2. Importantly, iPSC lines that pass these tests still vary in their capacity to produce different differentiated cell types2. This has made it difficult to establish which iPSC derivation protocols, donor cell sources or selection methods are most useful for different applications.
The most stringent test of whether a stem cell line has sufficient developmental potential to generate all tissues required for survival of an organism (termed full pluripotency) is tetraploid embryo complementation (TEC)3-5. Technically, TEC involves electrofusion of two-cell embryos to generate tetraploid (4n) one-cell embryos that can be cultured in vitro to the blastocyst stage6. Diploid (2n) pluripotent stem cells (e.g. ESCs or iPSCs) are then injected into the blastocoel cavity of the tetraploid blastocyst and transferred to a recipient female for gestation (see Figure 1). The tetraploid component of the complemented embryo contributes almost exclusively to the extraembryonic tissues (placenta, yolk sac), whereas the diploid cells constitute the embryo proper, resulting in a fetus derived entirely from the injected stem cell line.
Recently, we reported the derivation of iPSC lines that reproducibly generate adult mice via TEC1. These iPSC lines give rise to viable pups with efficiencies of 5-13%, which is comparable to ESCs3,4,7 and higher than that reported for most other iPSC lines8-12. These reports show that direct reprogramming can produce fully pluripotent iPSCs that match ESCs in their developmental potential and efficiency of generating pups in TEC tests. At present, it is not clear what distinguishes between fully pluripotent iPSCs and less potent lines13-15. Nor is it clear which reprogramming methods will produce these lines with the highest efficiency. Here we describe one method that produces fully pluripotent iPSCs and "all- iPSC" mice, which may be helpful for investigators wishing to compare the pluripotency of iPSC lines or establish the equivalence of different reprogramming methods.

Generating induced pluripotent stem cell (iPSC) lines produces lines of differing developmental potential even when they pass standard tests for pluripotency. Here we describe a protocol to produce mice derived entirely from iPSCs, which defines the iPSC lines as possessing full pluripotency1.

جيل من الفئران المستمدة من الخلايا الجذعية المحفزة المستحثة

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics

The liver is a complex and heterogenous tissue responsible for carrying out many critical physiological functions, such as the maintenance of energy homeostasis and the metabolism of xenobiotics, among others. These tasks are performed through tight coordination between hepatic parenchymal and non-parenchymal cells. Additionally, various metabolic activities are confined to specific areas of the hepatic lobule-a phenomenon called liver zonation. Recent advances in single-cell sequencing technologies have empowered researchers to investigate tissue heterogeneity at a single-cell resolution. In many complex tissues, including the liver, harsh enzymatic and/or mechanical dissociation protocols can negatively affect the viability or the quality of the single-cell suspensions needed to comprehensively characterize this organ in health and disease.
This paper describes a robust and reproducible protocol for isolating nuclei from frozen, archived liver tissues. This method yields high-quality nuclei that are compatible with downstream, single-cell omics approaches, including single-nucleus RNA-seq, assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq), as well as multimodal omics (joint RNA-seq and ATAC-seq). This method has been successfully used for the isolation of nuclei from healthy and diseased human, mouse, and non-human primate frozen liver samples. This approach allows the unbiased isolation of all the major cell types in the liver and, therefore, offers a robust methodology for studying the liver at the single-cell resolution.

Here, we present a protocol for isolating nuclei from flash-frozen, archived liver tissues for single-nucleus RNA-seq, ATAC-seq, and joint multiomics (RNA-seq and ATAC-seq).

عزل النوى من أنسجة الكبد المجمدة بالفلاش من أجل Multiomics أحادية الخلية

The liver is a complex and heterogenous tissue responsible for carrying out many critical physiological functions, such as the maintenance of energy ...

Single-Cell Suspension Preparation from Nile Tilapia Intestine for Single-Cell Sequencing

Nile tilapia is one of the most commonly cultured freshwater fish species worldwide and is a widely used research model for aquaculture fish studies. The preparation of high-quality single-cell suspensions is essential for single-cell level studies such as single-cell RNA or genome sequencing. However, there is no ready-to-use protocol for aquaculture fish species, particularly for the intestine of tilapia. The effective dissociation enzymes vary depending on the tissue type. Therefore, optimizing the tissue dissociation protocol by selecting the appropriate enzyme or enzyme combination to obtain enough viable cells with minimum damage is essential. This study illustrates an optimized protocol to prepare a high-quality single-cell suspension from Nile tilapia intestine with an enzyme combination of collagenase/dispase. This combination is highly effective for dissociation with the utilization of bovine serum albumin and DNase to reduce cell aggregation after digestion. The cell output satisfies the requirements for single-cell sequencing, with 90% cell viability and a high cell concentration. This protocol can also be modified to prepare a single-cell suspension from the intestines of other fish species. This research provides an efficient reference protocol and reduces the need for additional trials in the preparation of single-cell suspensions for aquaculture fish species.

Here, we demonstrate the preparation of a high-quality single-cell suspension of tilapia intestine for single-cell sequencing.

تحضير معلق أحادي الخلية من أمعاء البلطي النيلي لتسلسل الخلية الواحدة

Nile tilapia is one of the most commonly cultured freshwater fish species worldwide and is a widely used research model for aquaculture fish studies. The ...

عزل شبه آلي للأنسجة الدهنية البيضاء للفأر

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

The in vitro study of white, brown, and beige adipocyte differentiation enables the investigation of cell-autonomous functions of adipocytes and their mechanisms. Immortalized white preadipocyte cell lines are publicly available and widely used. However, the emergence of beige adipocytes in white adipose tissue in response to external cues is difficult to recapitulate to the full extent using publicly available white adipocyte cell lines. Isolation of the stromal vascular fraction (SVF) from murine adipose tissue is commonly executed to obtain primary preadipocytes and perform adipocyte differentiation. However, mincing and collagenase digestion of adipose tissue by hand can result in experimental variation and is prone to contamination. Here, we present a modified semi-automated protocol that utilizes a tissue dissociator for collagenase digestion to achieve easier isolation of the SVF, with the aim of reducing experimental variation, reducing contamination, and increasing reproducibility. The obtained preadipocytes and differentiated adipocytes can be used for functional and mechanistic analyses.

تسليط الضوء على المؤلف: عزل شبه آلي لجزء الأوعية الدموية اللحمية من الأنسجة الدهنية البيضاء الفئران باستخدام مفكك الأنسجة  

This protocol describes the semi-automated isolation of the stromal vascular fraction (SVF) from murine adipose tissue to obtain preadipocytes and achieve adipocyte differentiation in vitro. Using a tissue dissociator for collagenase digestion reduces experimental variation and increases reproducibility.

عزل شبه آلي للجزء الوعائي اللحمي من الأنسجة الدهنية البيضاء باستخدام مفكك الأنسجة

The in vitro study of white, brown, and beige adipocyte differentiation enables the investigation of cell-autonomous functions of adipocytes and their ...

هضم الخلايا من خطوتين للشريان السباتي للفأر

Preparation of a Single-Cell Suspension from Mouse Carotid Arteries for Single-Cell Sequencing

Carotid arteries are major blood vessels in the neck that supply blood and oxygen to the brain, but carotid stenosis occurs when carotid arteries are clogged by plaque. Revealing the cellular composition of the carotid artery at the single-cell level is essential for treating carotid atherosclerosis. However, there is no ready-to-use protocol for the preparation of single-cell suspensions from carotid arteries. To obtain a suitable protocol for the dissociation of normal carotid arteries at the single-cell level with less damage to cells, we designed a two-step digestion method by integrating the digestion process of collagenase/DNase and trypsin. Acridine orange/propidium iodide (AO/PI) dual-fluorescence counting was used to detect cell viability and concentration, and it was found that the single-cell suspension satisfied the requirements for single-cell sequencing, with the viability of cells over 85% and a high cell concentration. After single-cell data processing, a median of ~2500 transcripts per cell were detected in each carotid artery cell. Notably, a variety of cell types of the normal carotid artery, including vascular smooth muscle cells (VSMCs), fibroblasts, endothelial cells (ECs), and macrophages and dendritic cells (M&#966;/DCs), were concurrently detectable. This protocol may be applied to prepare a single-cell suspension of blood vessels from other tissues with appropriate modifications.

تسليط الضوء على المؤلف: تفكك أنسجة الأوعية الدموية واستكشاف المجموعات الفرعية أحادية الخلية للعلاج الموجه 

Here we describe a two-step cell digestion protocol for preparing a single-cell suspension of mouse carotid arteries.

تحضير معلق أحادي الخلية من الشرايين السباتية للفأر لتسلسل الخلية الواحدة

Carotid arteries are major blood vessels in the neck that supply blood and oxygen to the brain, but carotid stenosis occurs when carotid arteries are ...