אנו מודים עמיתינו ב-BSI RIKEN עבור תומך חביב שלהם עצות פורה במהלך עבודה זו. ברצוני להביע הערכה כנה שלי פרופ Shigemi מורי, פרופסור G בור Juh SZ, בני הזוג. Kiyoji Matsuyama, קתלין AK kesi ו RP ד Dobolyi אשר יעץ לי ללמוד שיטה microdialysis במהלך לתואר שני באוניברסיטת בוגר ללימודים מתקדמים, יפן. לבסוף, אנו מודים למר כריס Sarnecky וד&quot;ר Akitsugu Takasu עבור תומך שלהם הקלטת וידאו.

שלב 1. הכנת ההשתלה בדיקה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הרדמה (לא רואים בסרט) הרדימי כל ציפור עם זריקה interperitoneal של קטמין (12.5 מ&quot;ג / ק&quot;ג משקל גוף) ו xylazine (25 מ&quot;ג / ק&quot;ג) תערובת לייצר על רמה קבועה של הרדמה כירורגית. </li><li style=";text-align:right;direction:rtl"> תקן ראש למסגרת stereotaxic השתמש Xylocaine (TM) ג&#39;לי להפחית כאבים מקומיים של לחץ ברים &quot;האוזן. ראש של ציפור קבוע במנגנון stereotaxic המורכב ברים האוזן בעל מקור שמכיל את הראש בזווית של ° 45. במהלך הניתוח stereotaxic להשתלה, לחמם את הגוף של הציפור על ידי כרית כירורגית עם שליטה תרמית (חום = 34 ~ 36 ° C). </li><li style=";text-align:right;direction:rtl"> לחטא את אזור הניתוח לחטא את שדה הניתוח עם isodine (TM) לפני הניתוח. </li></ol> שלב 2. Stereotaxic מיצוב <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> נקודת התייחסות יישור יישר בדיקה דמה (זכוכית micropipette) על למבדה (Y נקודה) ו-Y סימן נקודה עם פיפטה צבע מלא. נותרת dextran-TMR המחקר נותב האחרון שימש הווידאו. </li><li style=";text-align:right;direction:rtl"> חתך של העור הסר את העור ולחשוף את פני השטח של הגולגולת. לאחר מכן, לנקות את פני השטח של הגולגולת עם צמר גפן. </li><li style=";text-align:right;direction:rtl"> המיקום של צינורית דמה (בכיוון אופקי) הזזת צינורית דמה לאתר היעד (AP ציר הרוחב ותיאום) </li><li style=";text-align:right;direction:rtl"> Craniotomy קטן חתך של מאטר הדורה. השתמש מלקחיים בסדר craniotomy ומחט הזרקת קנס (30 G) עבור חתכים של מאטר הדורה. לאחר מכן, לנקות את פני השטח של הגולגולת. </li><li style=";text-align:right;direction:rtl"> המיקום הסופי (ההשתלה) של צינורית דמה אתר את קצה של צינורית מדריך ב 0.30 מ&quot;מ מעל microdialysis בדיקה, נועד אזור היעד. לבסוף, לנקות את פני השטח של הגולגולת שוב, כי משטח נקי ויבש נדרש מליטה קשה. </li></ol> שלב 3. Pretreatment של פני הגולגולת (הליך דומה &quot;שיקום מילוי ישיר&quot; של המוצר שיניים יועסק). <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> פריימר טיפול <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מערבבים PRIMAR נוזלי עם B על צלחת. </li><li style=";text-align:right;direction:rtl"> החל את התערובת על פני השטח של הגולגולת עם מברשת קטנה. </li><li style=";text-align:right;direction:rtl"> השאירו למשך 30 שניות. </li><li style=";text-align:right;direction:rtl"> יבש עם זרם אוויר עדין. (לא לשטוף). </li></ol></li><li style=";text-align:right;direction:rtl"> יישום של סוכן מליטה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> החל הסוכן מליטה על פני הגולגולת עם מברשת קטנה. </li><li style=";text-align:right;direction:rtl"> החל זרם אוויר עדין. (לא לשטוף). </li><li style=";text-align:right;direction:rtl"> אור תרופה למשך 20 שניות. </li></ol></li></ol> שלב 4. הרכבה שרף מרוכב <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הר שרף עם plugger רגיל אם plugger רגיל אינו זמין, מרית מיקרו יכול לשמש במקום. </li><li style=";text-align:right;direction:rtl"> אור ריפוי </li><li style=";text-align:right;direction:rtl"> מצטבר הרכבה מצטבר הרכבה מומלץ להבטיח תגובה מלאה של שרף. עבור הרכבה מצטבר, להחיל סוכן מליטה לפני כל הרכבה. </li><li style=";text-align:right;direction:rtl"> עובש שרף מרוכבים אור ריפוי עבור התקשות סופית. </li><li style=";text-align:right;direction:rtl"> סוף הניתוח </li></ol> נקו את אזור הניתוח, חל אנטיביוטי (משחה גנטמיצין), ולהסיר את החיה מהאש stereotaxic. לאחר מכן, לשמור על בעלי החיים בתא תרמו שבשליטת (טמפרטורת = 33-35 ° C) להתאוששות לאחר הרדמה. כפי שניתן לראות בסרטון, התנהגות לא נורמלית נצפתה כל החיות למחרת לאחר הניתוח.

<ol>
	<li>Adell, A., Artigas, F., Boulton, A., Baker, G., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+Brain+microdialysis:+Principles+and+applications.">In vivo Brain microdialysis: Principles and applications.</a> In Vivo Neuromethods: Neuromethods. 32, 325-357 (1998).</li><li>Kendrick, K., Robinson, T., Justice, J., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microdialysis+in+large+unrestrained+animals:+neuroendocrine+and+behavioral+studies+of+acetylcholine,+amino+acid,+monoamine+and+neuropeptide+release+in+the+sheep.">Microdialysis in large unrestrained animals: neuroendocrine and behavioral studies of acetylcholine, amino acid, monoamine and neuropeptide release in the sheep.</a> Microdialysis in the Neuroscience. 7, 327-348 (1991).</li><li>Oakley, B., Schafer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Experimental Neurobiology: A laboratory Manual. , (1978).</li><li>Okumura, T., Yamashita, Y., Okanoya, K., Tani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+sensori-motor+nucleus+NIf+in+generation+of+complex+syntactical+song+in+the+Bengalese+Finch+I.+A+Biological+study.">Function of the sensori-motor nucleus NIf in generation of complex syntactical song in the Bengalese Finch I. A Biological study.</a> Soc. Neurosci Abst. 33, (2007).</li><li>Boulton, A., Baker, G., Robinson, T., Bateson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relation+of+brain+electrical+activity+to+behavior.">The relation of brain electrical activity to behavior.</a> In Vivo Neuromethods: Neuromethods. 32, 325-357 (1998).</li><li>Yamashita, Y., Okumura, T., Okanoya, K., Tani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+sensori-motor+nucleus+NIf+in+generation+of+complex+syntactical+song+in+the+Bengalese+finch.">Function of the sensori-motor nucleus NIf in generation of complex syntactical song in the Bengalese finch.</a> , 101-104 (2007).</li></ol>

הניסויים נערכו בהתאם הנחיות לניסויים בבעלי חיים של המוח המדע במכון, RIKEN (RIKEN-BSI) ובעלי חיים המכון של ועדת האתיקה אישרה אותם.

בהשוואה יישום קונבנציונאלי שרפים עצמי ריפוי אקריליק, הציג את הטכניקה יש מספר יתרונות חשובים: 1. ישיר מליטה: אקריליק ריפוי עצמי יכול להיות מצורף העצם למשך זמן מה אבל זה לא לדבוק העצם. לכן, ברגים מעגן נדרשים לתקן את בסיס הרכבה (ראה איור 2 א). לעומת זאת...

1A צילום מראה בדיקה microdialysis (א), אגוז כובע (ב) ו צינורית מדריך (ג). בתמונה זו, קנית את המדריך מוקף שרף שיניים הדוגלת הגולגולת ישירות. תמונה ב &#39;מראה ציפור במהלך זלוף סמים. בחלק העליון של הראש, יש בדיקה microdialysis שמחובר לצינור תפרון בסדר. ברים סולם = 1 ס&quot;מ ו 2 ס&quot;מ עבור B, בהתאמה. <img src="http://www.jove.com/files/249_2008_8_26_3/249_Figure-01.png" /> איור 1 - תצלומים של בדיקה microdialysis (א) ו ציפור במהלך זלוף סמים (ב)

application of light-cured dental adhesive resin for mounting electrodes or microdialysis probes in chronic experiments

בניסויים הקלטה כרונית, ריפוי עצמי שרף אקרילי שיניים שימשו בסיס הרכבה של אלקטרודות או microdialysis-בדיקות. מאז אקריליק אלה אינם האג&quot;ח עד העצם, ברגים שימשו עוגנים. עם זאת, בחיות מעבדה קטנות כמו פינקים או עכבר, הגולגלות שלהם הם שבירים מאוד לא מצליח להחזיק את העוגנים. בדו&quot;ח זה, אנו מציעים יישום חדש של אור ריפוי שיניים שרפים על בסיס הרכבה של אלקטרודות או בדיקות microdialysis בניסויים כרונית. חומר זה מאפשר התקשרות ישירה הגולגולת. לכן, ברגים עוגן לא נדרשים בתחום כירורגית ניתן להפחית במידה ניכרת. ניסיון העבר מראה כי ההשפעה מליטה שומרת יותר מ 2 חודשים. חלון של שרף קונבנציונלי של זמן כאשר החומרים הם וגמיש ו עביד הוא כמה דקות. עם זאת, חלון של זמן עבודה עבור אלה הדבקים שיניים משמעותית רחבה מתכווננת.

Watch this Scientific Journal Video about Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments at JoVE.com

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

בדו&quot;ח זה, אנו מציעים יישום חדש של אור ריפוי שיניים שרפים על בסיס הרכבה של אלקטרודות או בדיקות microdialysis בניסויים כרונית. חומר זה מאפשר התקשרות ישירה הגולגולת.

יישום של שרף אור נרפא, דבק שיניים אלקטרודות הרכבה או בדיקות Microdialysis בניסויים כרונית

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

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Research

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Biology

15.3K Views.  RIKEN. בדו"ח זה, אנו מציעים יישום חדש של אור ריפוי שיניים שרפים על בסיס הרכבה של אלקטרודות או בדיקות microdialysis בניסויים כרונית. חומר זה מאפשר התקשרות ישירה הגולגולת.

Video: Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

Preparation of Artificial Bilayers for Electrophysiology Experiments

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. These bilayers can be used for many different studies, such as the characterization of membrane-active peptides, the reconstitution of ion channels or investigations on how changes in lipid bilayer properties alter the function of bilayer-spanning channels.  Here, we show how to form a planar bilayer and how to isolate small patches from the bilayer, and in a second video  will also demonstrate a procedure for using gramicidin channels to determine changes in lipid bilayer elastic properties.   We also demonstrate the individual steps needed to prepare the bilayer chamber, the electrodes and how to test that the bilayer is suitable for single-channel measurements.

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. Here, we demonstrate the individual steps needed to prepare the bilayer chamber, the electrodes and how to test that the bilayer is suitable for single-channel measurements.

הכנת Bilayers מלאכותית לניסויים Electrophysiology

Planar lipid bilayers, also called artificial lipid bilayers, allow you to study ion-conducting channels in a well-defined environment. These bilayers can ...

Fabrication of Amperometric Electrodes

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we describe the techniques needed to generate low noise (&lt;0.5 pA) electrodes.  The techniques have been modified from published descriptions by previous researchers (1,2).  Electrodes are made by preparing carbon fibers and threading them individually into each capillary tube by using a vacuum with a filter to aspirate the fiber.  Next, the capillary tube with fiber is pulled by an electrode puller, creating two halves, each with a fine-pointed tip.  The electrodes are dipped in hot, liquid epoxy mixed with hardener to create an epoxy-glass seal.  Lastly, the electrodes are placed in an oven to cure the epoxy.  Careful handling of the electrodes is critical to ensure that they are made consistently and without damage.  This protocol shows how to fabricate and cut amperometric electrodes for recording from single cells.

This protocol describes how to generate carbon fiber electrodes. The electrodes are subsequently used to detect catecholamine release from vesicles with carbon fiber amperometry.

המצאה של אלקטרודות Amperometric

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we ...

Robust 3D DNA FISH Using Directly Labeled Probes

3D DNA FISH has become a major tool for analyzing three-dimensional organization of the nucleus, and several variations of the technique have been published. In this article we describe a protocol which has been optimized for robustness, reproducibility, and ease of use. Brightly fluorescent directly labeled probes are generated by nick-translation with amino-allyldUTP followed by chemical coupling of the dye. 3D DNA FISH is performed using a freeze-thaw step for cell permeabilization and a heating step for simultaneous denaturation of probe and nuclear DNA. The protocol is applicable to a range of cell types and a variety of probes (BACs, plasmids, fosmids, or Whole Chromosome Paints) and allows for high-throughput automated imaging. With this method we routinely investigate nuclear localization of up to three chromosomal regions.

We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.

דגי ה-DNA 3D חזקים באמצעות בדיקות שכותרתו ישירות

3D DNA FISH has become a major tool for  analyzing three-dimensional organization of the nucleus, and several variations  of the technique have been ...

Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish

Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is particularly suited for long-term time-lapse imaging over many hours up to several days. However, an appropriate sample mounting strategy is needed that offers both confinement and normal development of the sample. Multilayer mounting, a new embedding technique using low-concentration agarose in optically clear tubes, now overcomes this limitation and unleashes the full potential of light sheet microscopy for real-time developmental biology.

The development of zebrafish can be followed over days with light sheet microscopy when embryos are embedded in optically clear polymer tubes with low-concentration agarose.

רב שכבתי וההרכבה למיקרוסקופיה גיליון אור לטווח ארוך של דג הזברה

Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is ...

Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse incisor provides a model for these processes. This remarkable organ grows continuously throughout the animal&#39;s life and generates all the necessary cell types from active pools of adult stem cells housed in the labial (toward the lip) and lingual (toward the tongue) cervical loop (CL) regions. Only the dental stem cells from the labial CL give rise to ameloblasts that generate enamel, the outer covering of teeth, on the labial surface. This asymmetric enamel formation allows abrasion at the incisor tip, and progenitors and stem cells in the proximal incisor ensure that the dental tissues are constantly replenished. The ability to isolate and grow these progenitor or stem cells in vitro allows their expansion and opens doors to numerous experiments not achievable in vivo, such as high throughput testing of potential stem cell regulatory factors. Here, we describe and demonstrate a reliable and consistent method to culture cells from the labial CL of the mouse incisor.

The continuously growing mouse incisor provides a model for studying renewal of dental tissues from dental epithelial stem cells (DESCs). A robust system for consistently and reliably obtaining these cells from the incisor and expanding them in vitro is reported here.

בידוד והתרבות של תאי גזע השיניים אפיתל ממבוגרי העכבר הארקטיק

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse ...

The Assembly and Application of 'Shear Rings': A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as well as pathological changes in endothelial phenotype and gene expression. In particular, maladaptive effects of periodic, unidirectional flow induced shear stress can trigger a variety of effects on several vascular cell types, particularly endothelial cells. While by now endothelial cells from diverse anatomic origins have been cultured, in-depth analyses of their responses to fluid shear have been hampered by the relative complexity of shear models (e.g., parallel plate flow chamber, cone and plate flow model). While these all represent excellent approaches, such models are technically complicated and suffer from drawbacks including relatively lengthy and complex setup time, low surface areas, requirements for pumps and pressurization often requiring sealants and gaskets, creating challenges to both maintenance of sterility and an inability to run multiple experiments. However, if higher throughput models of flow and shear were available, greater progress on vascular endothelial shear responses, particularly periodic shear research at the molecular level, might be more rapidly advanced. Here, we describe the construction and use of shear rings: a novel, simple-to-assemble, and inexpensive tissue culture model with a relatively large surface area that easily allows for a high number of experimental replicates in unidirectional, periodic shear stress studies on endothelial cells.

The Assembly and Application of &#039;Shear Rings&#039;: A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

Different levels and patterns of fluid shear are known to modulate endothelial gene expression, phenotype and susceptibility to disease. We discuss the assembly and use of 'shear rings': a model that produces unidirectional, periodic shear stress patterns. Shear rings are simple to assemble, economical and can produce high cell yields.

האסיפה והיישום של &#39;טבעות Shear&#39;: מודל האנדותל חדשני Orbital, חד-כיוונית תקופתית נוזלי זרימת מתח שאר

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as ...

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells

Nitric Oxide (NO&#8226;) is a small radical, which mediates multiple important cellular functions in mammals, bacteria and plants. Despite the existence of a large number of methods for detecting NO&#8226; in vivo and in vitro, the real-time monitoring of NO&#8226; at the single-cell level is very challenging. The physiological or pathological effects of NO&#8226; are determined by the actual concentration and dwell time of this radical. Accordingly, methods that allow the single-cell detection of NO&#8226; are highly desirable. Recently, we expanded the pallet of NO&#8226; indicators by introducing single fluorescent protein-based genetically encoded nitric oxide (NO&#8226;) probes (geNOps) that directly respond to cellular NO&#8226; fluctuations and, hence, addresses this need. Here we demonstrate the usage of geNOps to assess intracellular NO&#8226; signals in response to two different chemical NO&#8226;-liberating molecules. Our results also confirm that freshly prepared 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7) has a much higher potential to evoke change in intracellular NO&#8226; levels as compared with the inorganic NO&#8226; donor sodium nitroprusside (SNP). Furthermore, dual-color live-cell imaging using the green geNOps (G-geNOp) and the chemical Ca2+ indicator fura-2 was performed to visualize the tight regulation of Ca2+-dependent NO&#8226; formation in single endothelial cells. These representative experiments demonstrate that geNOps are suitable tools to investigate the real-time generation and degradation of single-cell NO&#8226; signals in diverse experimental setups.

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&amp;#8226;) Probes, the geNOps, for Real-time Imaging of NO&amp;#8226; Signals in Single Cells

This manuscript presents protocols for the application of novel genetically encoded nitric oxide (NO&#8226;) probes (geNOps) to monitor single cell NO&#8226; fluctuations in real-time using fluorescence microscopy. The Ca2+-triggered NO&#8226; formation on the level of individual endothelial cells was visualized by combining geNOps with a chemical Ca2+ sensor.

יישום של תחמוצת החנקן פלורסנט מקודדים גנטית (NO •) בדיקות, את geNOps, הדמיה בזמן אמת של אותות NO • ב תאים בודדים

Nitric Oxide (NO&#8226;) is a small radical, which mediates multiple important cellular functions in mammals, bacteria and plants. Despite the existence ...

Inducing Apical Periodontitis in Mice

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical periodontitis (AP). Apical periodontitis is a common pathology of an inflammatory nature in the periodontal tissues surrounding the tooth root. In order to better understand the nature and mechanism of this pathology it is advantageous to perform the procedure in mice. The induction of this odontogenic inflammation is achieved by drilling into the mouse tooth until the dental pulp is exposed. Next, the tooth pulp remains exposed to be contaminated by the natural oral flora over time, causing apical periodontitis. After this time period, the animal is sacrificed, and the tooth and the jaw bone can be analyzed in various ways. Typical analyses include micro-CT imaging (to evaluate bone resorption), histological staining, immunohistochemistry, and RNA expression. This protocol is useful for research in the field of oral biology to better understand this inflammatory process in an in vivo experimental setting with uniform conditions. The procedure requires a careful handling of the mice and the isolated jaw, and a visual demonstration of the technique is useful. All technical aspects of the procedures leading to induced apical periodontitis and its characterization in a mouse model are demonstrated.

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse&#39;s tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

לגרימת חניכיים הרשמי בעכברים

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical ...

Systematic Assessment of Mammalian Skull Specimens for Dental and Temporomandibular Joint Pathology

Museum skull specimens represent a non-invasive, informative, and readily available means to study temporomandibular joint (TMJ) lesions, dental pathology, and anatomic variations in many mammalian species. Studying the teeth and jaws of an array of species can present a challenge requiring attention to detail and understanding of a species&#39; normal anatomy. In the present article, a systematic and precise protocol for examining skull specimens is discussed that has been applied to a variety of mammals to define characteristic diseases in the oromaxillofacial region. The procedure outlined is simultaneously precise, repeatable, and adaptable to the highly differing skull and tooth shapes and anatomy across species. Specifically, specimens are examined for missing teeth, periodontal disease, endodontal disease, TMJ pathology, and anatomical variations. Results gleaned from research on museum specimens may reflect the natural history, health, and disease status of individuals and species. Furthermore, these data can inform ecological and conservation research efforts, as well as the care of captive individuals.

The present protocol describes the techniques for the systematic assessment of skull specimens to characterize anatomical and developmental variations and abnormalities of the teeth, periodontal disease, endodontal disease, and temporomandibular joint pathology.

הערכה שיטתית של דגימות גולגולת יונקים לפתולוגיה דנטלית וטמפורומנדיבולרית של המפרקים

Museum skull specimens represent a non-invasive, informative, and readily available means to study temporomandibular joint (TMJ) lesions, dental ...

ביסוס התרבית הראשונית של תאי גזע במוך השן

Primary Culture of Dental Pulp Stem Cells

The human dental pulp represents a promising multipotent stem cell reservoir with pre-eminent regenerative competence that can be harvested from an extracted tooth. The neural crest-derived ecto-mesenchymal origin of dental pulp stem cells (DPSCs) bestows a high degree of plasticity that owes to its multifaceted benefits in tissue repair and regeneration. There are various practical ways of harvesting, maintaining, and proliferating adult stem cells being investigated for their use in regenerative medicine. In this work, we demonstrate the establishment of a primary mesenchymal stem cell culture from dental tissue by the explant culture method. The isolated cells were spindle-shaped and adhered to the plastic surface of the culture plate. The phenotypic characterization of these stem cells showed positive expression of the international society of cell therapy (ISCT)-recommended cell surface markers for MSC, such as CD90, CD73, and CD105. Further, negligible expression of hematopoietic (CD45) and endothelial markers (CD34), and less than 2% expression of HLA-DR markers, confirmed the homogeneity and purity of the DPSC cultures. We further illustrated their multipotency based on differentiation to adipogenic, osteogenic, and chondrogenic lineages. We also induced these cells to differentiate into hepatic-like and neuronal-like cells by adding corresponding stimulation media. This optimized protocol will aid in the cultivation of a highly expandable population of mesenchymal stem cells to be utilized in the laboratory or for preclinical studies. Similar protocols can be incorporated into clinical setups for practicing DPSC-based treatments.

מחבר זרקור: קידום התחדשות רקמות ומידול מחלות עם תאי גזע במוך השן

This article provides a stepwise guide to establish a primary culture of dental pulps stem cells using the explant culture method and characterization of these cells based on ICSCRT guidelines. The cells isolated by this protocol can be considered as mesenchymal stem cells for further applications.

תרבית ראשונית של תאי גזע במוך השן

The human dental pulp represents a promising multipotent stem cell reservoir with pre-eminent regenerative competence that can be harvested from an ...