Die Autoren bedanken sich bei Dr. Ana Cotrim für technische Beratung und Dr. Monzur für Geräte Murshed. Diese Arbeit wurde zum Teil durch die Canadian Institutes for Health Research finanziert.

1. Verfahrensbeschreibung <ol><li> Subkutan zu injizieren 1 ul / g Körpergewicht von 0,5 mg / ml Atropinsulfat-Monohydrat (Kat. Nr. 11330, Fluka, USA) in jede Maus an den Speicheldrüsen Sekret von der Störung der Bewegung der Infusionen zu verhindern. </li><li> Anesthetize jeder Maus mit 1 ul / g Körpergewicht von 100 mg / ml Ketamin und 20 mg / ml Xylazin (Phoenix Scientific, AR, USA) injiziert intraperitoneale (IP) </li><li> Legen Sie die Maus auf eine maßgeschneiderte Kunststoff-Plattform in der Bauchlage: Die oberen Schneidezähne sind auf einem Metalldraht gesperrt, und die unteren Schneidezähne sind auf einer elastischen Schnur eingehakt um in den Besitz der Mund offen. </li><li> Verwenden Sie eine Insulin-Spritze mit einer 29-Gauge-Nadel (Cat # 600145, Tyco Healthcare, MA, USA) in einem 0,58 mm Durchmesser Polyethylen Rohr (Cat # 427410, Becton Dickinson, MD, USA; Abb. 2) platziert. Die Polyethylen-Rohr eingesetzt ist 5mm über der Spitze einer 29-Gauge-Nadel. Die Nadel ist wie ein starrer Leitfaden für die Polyethylen-Rohr in die Wharton-Gang bewegen verwendet. Die Polyethylen-Rohr wird 3-5 mm in den Kanal eingeschoben. Beachten Sie, dass nur die Polyethylen-Rohr der Speicheldrüse Kanal gelangt, während der Nadel bleibt außerhalb des Kanals. Inject 50ul der gefilterten 0,4% Trypanblau Färbelösung (Cat # 15250, Gibco, USA) in die submandibularis über die Wharton-Gang, der direkt unterhalb der Zunge liegt. Die Infusion wird ständig mit positiven Druck auf die Spritze Kolben geliefert, der Finger bleibt auf dem Kolben auch nach all der Infusion hat, um die Drüse geliefert worden, um einen Rückfluss der Infusion intraoral zu verhindern. Der Trypanblau-Lösung ist in dem Video als Farbstoff, um die Technik zu demonstrieren. </li><li> N ach der Injektion der Trypanblau-Lösung wurde die Maus vor der Dissektion eingeschläfert. Die Speicheldrüse wird operativ in Ordnung ausgesetzt, um festzustellen, wie viel von der Infusionen tatsächlich in der Drüse blieb. </li></ol> 2. Repräsentative Ergebnisse: Nach der Infusion injiziert wurden, war die Maus seziert, um operativ Setzen Sie das post-kanülierten Speicheldrüse. Wie auf Bild-2 gezeigt, stellt die dunkle Färbung der Wharton-Gang Beweise, dass die Trypanblau-Lösung wurde erfolgreich infundiert. <img src="/files/ftp_upload/3074/3074fig1.jpg" alt="Abbildung 1" /> Abbildung 1. Kanülierung der Maus submandibularis. <img src="/files/ftp_upload/3074/3074fig2.jpg" alt="Abbildung 2" /> Abbildung 2. Polyethylen Rohr ummantelt um die Spitze eines 29-Gauge-Nadel. Die Kanüle wird als starre Führung des Polyethylen-Rohr in die Wharton-Gang bewegen verwendet. Die Polyethylen-Rohr (kanülierten) 3-5 mm in den Kanal eingeschoben. Beachten Sie, dass nur die Polyethylen-Rohr der Speicheldrüse Kanal gelangt, während der Nadel bleibt außerhalb des Kanals.

<ol>
	<li>Fox, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+salivary+dysfunction.+Drugs+and+radiation.">Acquired salivary dysfunction. Drugs and radiation.</a> Ann N Y Acad Sci. 842, 132-137 (1998).</li><li>Bhide, S. A., Miah, A. B., Harrington, K. J., Newbold, K. L., Nutting, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation-induced+xerostomia:+pathophysiology,+prevention+and+treatment.">Radiation-induced xerostomia: pathophysiology, prevention and treatment.</a> Clin Oncol (R Coll Radiol). 21, 737-744 (2009).</li><li>Redman, R. S., Ball, W. D., Mezey, E., Key, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dispersed+donor+salivary+gland+cells+are+widely+distributed+in+the+recipient+gland+when+infused+up+the+ductal+tree.">Dispersed donor salivary gland cells are widely distributed in the recipient gland when infused up the ductal tree.</a> Biotech Histochem. 84, 253-260 (2009).</li></ol>

Alle Experimente wurden von der Universität Animal Care Committee genehmigt worden und wurden in Übereinstimmung mit den Richtlinien des Canadian Council on Animal Care.

Eine Schwierigkeit mit diesem Kanülierung Methode besteht darin, den Rückfluss der injizierten Lösung zu verhindern. Selbst wenn der Trypanblau-Lösung wurde vorübergehend in die Speicheldrüse erfolgreich migriert, kann der Speichel-Sekretion mit der Infusionen und verursachen Rückfluss der Trypanblau-Lösung aus der Drüse. Als Vorsichtsmaßnahme zusätzlich zur Verwendung Atropin zur Sekretion von Speichel hemmen, sollten die Polyethylenkanüle in die Wharton-Gang und der Kolben der Spritze für 60 Sekunden gehalten, nachdem die Infusionen hinterlegt haben schon bleiben. Damit können die meisten der infundierten Lösung innerhalb der Drüse bleiben. Üblicherweise ist die systemische Anwendung, nämlich die intravenöse Injektion (IV) von Drogen / Zellen, als einen klinischen Ansatz für Speicheldrüse Regeneration eingesetzt. Intravenös injiziert Infusionen wird durch das gesamte Kreislaufsystem reisen, so dass nur ein Teil der Drogen / Zellen, in die gezielte Drüsen zu migrieren. Im Vergleich zu dieser traditionellen Methode kann die lokale Anwendung, dass die Injektion von Drogen / Zellen über die Kanülierung Verfahren ist, wie eine direkte und effektive Art und Weise in die Wiederbelebung der Funktionen der Parotis, submandibularis und sublingualen Speicheldrüsen wahrgenommen werden. Deshalb kann diese Kanülierung Technik als ein zuverlässiges Modell für die regenerative Medizin von Maus Speicheldrüse verwendet werden, und als eine Perspektive, dieses Verfahren kann eine sinnvolle Methode für die klinische Anwendung sein. Es wäre eine große Freude, diese visuell-Artikel als ein effektives Werkzeug für Studenten und Forscher, die in weiteren Studien in der regenerativen Medizin der Speicheldrüse interessiert sind, verwendet werden.

<table border="1"><tbody><tr><td> Name des Reagenzes </td><td> Typ </td><td> Firma </td><td> Katalog-Nummer </td><td> Kommentare (optional) </td></tr><tr><td> Atropin </td><td> Medikament </td><td> Fluka </td><td> 11330 </td><td> Zur Inhibierung Speichel-Sekretion </td></tr><tr><td> Ketamin </td><td> Medikament </td><td> Phoenix Scientific </td><td></td><td> Beruhigung </td></tr><tr><td> Xylazin </td><td> Medikament </td><td> Phoenix Scientific </td><td></td><td></td></tr><tr><td> Polyethylen-Schlauch (0,58 mm Durchmesser) </td><td> Tube </td><td> Intramedic PE-10, Becton Dickinson Diagnostic Systems </td><td> 427410 </td><td> Um mehr als Spitze der Nadel einlegen, um Gewebeschäden beim Einführen in Wharton-Gang vermeiden. </td></tr><tr><td> Ultra Comfort Insulin-Spritze (mit einer 29G Nadel) </td><td> Spritze </td><td> Tyco Healthcare </td><td> 600145 </td><td></td></tr><tr><td> 4% Trypanblau </td><td> Fleck </td><td> Gibco </td><td> 15250 </td><td></td></tr></tbody></table>

cannulation of the mouse submandibular salivary gland via the wharton's duct

Schwere Speicheldrüse Unterfunktion ist häufig bei Patienten mit Sjögren-Syndrom und diejenigen, die therapeutischen Bestrahlung erhalten in ihrem Kopf und Hals Regionen zur Behandlung von Krebs gefunden. Beide Gruppen von Patienten Symptome wie Xerostomie (Mundtrockenheit), Dysphagie (Störungen der Kau-und Schluckstörungen), schwere Karies, verändertes Geschmacksempfinden, Mund-Rachen-Infektionen (Candida), Mukositis, Schmerzen und Beschwerden. . Ein innovativer Ansatz der regenerativen Medizin für die Behandlung von Speicheldrüsen hypo-Funktion ist in RS Redman, E Mezey et al 2009 spekuliert: Stammzellen können direkt durch Kanülierung werden in der Drüse abgelagert als wirksame Methode bei der Wiederbelebung der Funktionen der gestörten Orgel. Vermutlich werden die migrierten ausländischen Stammzellen in Drüsenzellen, die als Teil der Host-Funktion der Speicheldrüsen unterscheiden. Auch dieses ist Kanülierung Technik eine sinnvolle und effektive Methode für die klinische Gentransfer-Anwendung. Hier zeigen die Schritte bei der Durchführung der Kanülierung Verfahren auf der Maus Glandula Speicheldrüse über die Wharton-Gang (Abb. 1) beteiligt. C3H-Mäuse (Charles River, Montreal, QC, Canada) sind für dieses Experiment, das unter sauberen konventionellen Bedingungen wurden an der McGill University Tier Resource Center gehalten werden. Alle Experimente wurden von der Universität Animal Care Committee genehmigt worden und wurden in Übereinstimmung mit den Richtlinien des Canadian Council on Animal Care. Für dieses Experiment ist eine Trypanblau-Lösung in die Drüse durch die Öffnung des Wharton-Gang mit einer Insulin-Spritze mit einer 29-Gauge-Nadel eingehüllt in einem Polyethylen-Schlauch infundiert. Anschließend wird die Maus seziert, um zu zeigen, dass die Infusionen in die Drüse erfolgreich migriert.

Watch this Scientific Journal Video about Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct at JoVE.com

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Ein Protokoll für die Kanülierung der Maus Glandula Speicheldrüse über die Wharton-Gang beschrieben. Für dieses Experiment ist die Trypanblau-Lösung als Färber verwendet werden, um zu demonstrieren, wie diese Technik effektiv liefert Infusionen in die gezielte Drüse, und die Zuverlässigkeit dieses neuen Ansatzes als potenzielle klinische Arzneimittelentwicklung / Zelltherapie für die Regeneration der Speicheldrüsen vorschlagen.

Kanülierung der Maus Submandibular Speicheldrüsen über die Wharton-Gang

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

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Nanyang Technological University

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Medicine

25.9K Aufrufe.  McGill University. Ein Protokoll für die Kanülierung der Maus Glandula Speicheldrüse über die Wharton-Gang beschrieben. Für dieses Experiment ist die Trypanblau-Lösung als Färber verwendet werden, um zu demonstrieren, wie diese Technik effektiv liefert Infusionen in die gezielte Drüse, und die Zuverlässigkeit dieses neuen Ansatzes als potenzielle klinische Arzneimittelentwicklung / Zelltherapie für die Regeneration der Speicheldrüsen vorschlagen.

Serie: Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Permanent Cerebral Gefäßverschluss<em&gt; Über</em&gt; Double Blattschraube und Durchtrennung

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

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Medizin

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Technische Aspekte der Maus aortokavaler Fistel

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

Maus Nierentransplantation: Modelle der Transplantatabstoßung

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

This protocol describes an injury model involving the surgical ligation of the pancreatic duct in the adult mouse pancreas, resulting in severe injury that establishes an environment that allows beta cell neogenesis and proliferation. This model can be used as a tool to study mechanisms involved in beta cell formation.

Chirurgische Verletzung des Maus-Pankreas durch Ligatur des Pankreasganges als Modell für endokrine und exokrine Reprogrammierung und Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. Irreparable cell damage results in a chronic loss of salivary function that impairs basic oral activities, and increases the risk of oral infections and dental caries. Salivary hypofunction and its complications gravely impact a patient's comfort. Current symptomatic management of the condition is ineffective, and newer therapies to assuage the condition are needed. 
Salivary glands are exocrine glands, which expel their secretions into the mouth via excretory ducts. Cannulation of these ducts provides direct access to the glands. Retroductal delivery of a contrast agent to major salivary glands is a routine out-patient procedure for diagnostic imaging. Using a similar procedure, localized treatment of the glands is feasible. However, performing this technique in preclinical studies with small animals poses unique challenges. In this study we describe the technique of retroductal administration in rat submandibular glands, a procedure that was refined in Dr. Bruce Baum's laboratory (NIH)1, and lay out a procedure for local gland irradiation.

Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands.

Retroductal Submandibulardrüse Einträufeln und Lokalisierte fraktionierten Bestrahlung in einem Rattenmodell von Speichel- Unterfunktion

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

Pankreasgang-Infusion: Eine effektive und selektive Methode der Medikamenten- und Virusabgabe

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In these cases, questions remain regarding disease pathogenesis and effective interventions. An optimized technique that allows functional assessment of the salivary glands is invaluable for investigating exocrine gland biology, dysfunction, and therapeutics. Here, we present a step by step approach to performing pilocarpine stimulated saliva secretion, including tracheostomy and the dissection of the three major murine salivary glands. We also detail the appropriate murine head and neck anatomy accessed during these techniques. This approach is scalable, allowing for multiple mice to be processed simultaneously, thus improving the efficiency of the work flow. We aim to improve the reproducibility of these methods, each of which has further applications within the field. In addition to saliva collection, we discuss metrics for quantifying and normalizing functional capacity of these tissues. Representative data are included from submandibular glands with depressed salivary gland function 2 weeks following fractionated radiation (4 doses of 6.85 Gy).

We present a detailed approach to performing saliva collection, including murine tracheostomy and the isolation of three major salivary glands.

Murine Speichel funktionale Bewertung über Pilocarpin Stimulation folgenden fraktioniert Bestrahlung

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In ...

Radiation Treatment of Organotypic Cultures from Submandibular and Parotid Salivary Glands Models Key In Vivo Characteristics

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.

Using three-dimensional organotypic cultures to visualize morphology and functional markers of salivary glands may provide novel insights into the mechanisms of tissue damage following radiation. Described here is a protocol to section, culture, irradiate, stain, and image 50&#8211;90 &#956;m thick salivary gland sections prior to and following exposure to ionizing radiation.

Strahlenbehandlung von organischen Kulturen aus Submandibulären und Parotid-Salivarienmodels Schlüssel in Vivo-Leiten

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

Ein Mausmodell für chronische Pankreatitis mittels Gallengangs-TNBS-Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...