Name: retroductal submandibular gland instillation and localized fractionated irradiation in a rat model of salivary hypofunction
Uploaded: 2016-04-24T15:00:00.000+00:00
Duration: 7 min 44 s
Description: Watch this Scientific Journal Video about Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction at JoVE.com

We thank Dr. John Chang (Radiation Oncology, LSU Health Science Center) for assistance with radiation dose measurements. The study was supported by the American Cancer Society (Grant number: 116945-RSG-09-038-01-CCE), National Institute of Health (Grant number: R21CA173162) and the Feist-Weiller Cancer Center.

所有的程序是由路易斯安那州立大学健康，什里夫波特，动物护理和使用委员会批准，并分别按照照顾和使用实验动物的指导方针NIH。 1.鼠颌下腺涎腺插管<ol><li> 注射器管组件的研制 <ol><li>切PE10聚乙烯管的一个10cm的长度用手术刀。容纳食指和拇指之间的管道的两端。加热上述平缓火焰管的中间部分，并轻轻拉伸软化的管道通过拉双方加倍其长度。 </li><li>在切割用手术刀中间管以45°角拿到2套管-每一个锥形端。通过在0.5毫升的胰岛素注射器的29克针嵌合它扩大插管的非锥形端。 </li><li>取出套管，并绘制220微升盐溶液进入注射器。点击赶走气泡。适合cannul一个在针，推动注射器的柱塞以排出空气，并确保通过套管的溶液自由流动。音量调整到200微升。 </li></ol></li><li> 颌下腺导管的口腔内插管 注:高压灭菌仪器之前的程序，并在两者之间进行消毒在热珠灭菌程序。 <ol><li>权衡只SD大鼠，并分配氯胺酮的计算量（42毫克/千克）/赛拉嗪（8毫克/千克）/乙酰丙嗪（1.4毫克/千克）的混合在一个皮下注射器。 </li><li>通过抓住食指和一只手的拇指之间的尾巴的基部和滑另一方面在主体把握它抑制动物。其余部分的食指和中指沿头部两侧，同时容纳用拇指和其余手指躯干。 </li><li>注入在后肢肌肉组织的麻醉剂。确认通过脚趾捏和眼睑反射麻醉深度。敷眼润滑剂防止干燥而动物处于麻醉状态。 </li><li>放置一个专门设计的平台（ 图1）上的动物，和横杆上接合的上门牙。通过循环围绕下门牙的橡胶带和它锚定到平台拉下颌向下。 </li><li>通过无菌缝合通过舌头将其提起，以提高口底。夹住缝合用止血，并且将它传递横杆。 </li><li>扩大与定制的脸颊吊具（ 图1）的脸颊，并在解剖显微镜下找到在口底舌下乳头。 </li><li>掌握使用细腻镊子预制PE10管的锥形端。轻轻操纵套管的尖端插入舌下乳头乳腺导管口。通过线程它3确认导管的放置 - 5 mm产品到管道;确保它传递而没有任何阻碍。 </li></ol></li><li> 颌下腺Instillation <ol><li>注射阿托品（0.5毫克/千克）皮下的颈背，并等待10分钟在唾液分泌减少。 </li><li>固定在插管到导管孔用氰基丙烯酸酯（胶）的下降，并使其干燥。灌输通过在〜50微升/分钟的速度缓慢地按下针筒柱塞在压盖（200微升/腺）的溶液中。 </li><li>粉碎用止血钳管道，并小心地取出注射器。保留在管道中的管道30 - 60分钟，直到动物恢复意识。卸下将舌头缝合。 </li><li>动物转移到一个单独的笼子里，并用热灯恢复过程中保持温暖。不要离开无人看管的动物，直到它已恢复了意识，以保持胸骨靠着。 </li><li>动物是完全卧床后，在不受限制地访问食物和水动物饲养安置它。 </li></ol></li></ol> 2.局部分割照射颌下腺<ol><li>抑制动物如之前描述的，并用氯胺酮肌内给药（33毫克/千克）/赛拉嗪（6毫克/公斤）/乙酰丙嗪（1毫克/千克）混合在后肢麻痹。确认麻醉深度，并使用润滑剂涂抹到眼睛。 </li><li>广场上的直线加速器桌面动物仰卧，并通过倾斜头部延长它的脖子。准直辐射场3厘米（狭缝宽度），以从下颌骨到胸骨的顶部的下边界包围的区域。 </li><li>放置1厘米组织等效丸剂以上的区域中，并调整辐射源和丸剂至100cm的顶部之间的距离。 </li><li>照射使用线性加速器的6 MV光子束的动物（2.5 Gy）进行。从辐射源剂量率，字段大小和距离弹丸表面将决定的曝光时间。在设置当前，动物在〜1戈瑞/分钟的剂量率照射。 </li><li>重复曝光;总共8 2.5格雷/天天; 4天/周，其间有2天的间隔。保持动物的体温恢复过程中。动物转移到动物饲养后，它完全卧床。 </li><li>收集刺激下颌下腺唾液8周照射后，测量腺体功能14。由冷的4％多聚甲醛/磷酸盐缓冲盐水pH 7.2的的心脏灌注安乐死麻醉下的动物。消灭颌下腺组织学和免疫组化分析14。 </li></ol>

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The authors have nothing to disclose.

唾液腺经常接受的辐射剂量超出经历了头颈部肿瘤，颈淋巴结选修消融或地区血液系统恶性肿瘤放疗的病人组织恢复的门槛。尽管腺的流体分泌腺泡细胞终末分化，它们是矛盾的对辐射敏感。分泌功能辐射和不可逆转的破坏腺体结果的第一周内滴在慢性低口水输出。为了打击穷人腺体功能和随之而来的口腔干燥，保存或分泌功能的恢复是至关重要的。新的治疗方法，以防止或修复辐射的伤害正在?...

健康组织的侧支破坏占了许多癌症治疗的有害副作用。与该辐射场位于大唾液腺的一部分或全部都不可避免地破坏。因此，经历骨髓移植前头部和颈部癌，子宫颈癌淋巴瘤，或全身辐射放疗大多数患者遭受的辐射，唾液腺机能减退2-6的最常见和最 ​​持久的不利效果之一。 唾液腺的流体产生腺泡细胞对辐射十分敏感。损坏唾液腺导致唾液流，称为唾液腺机能减退的条件的急剧减少。在唾液流量慢性减少损害的关键口腔活动，如咀嚼，吞咽，语言和口味，但剧烈的疼痛，黏膜撕裂，吞咽困难，机会性感染病态的后遗症，和龋齿恶化一个病人的福祉和功能2,3。 由于放疗相关的唾液腺细胞的损失是不可逆的，有口干没有矫正治疗。专注于人工唾液替代品和药品prosecretory平息目前的症状治疗无效的长期救援6。虽然改进的辐射递送技术已经帮助减少的病症的严重程度，正常组织的毒性及其并发症留在癌症治疗中6,7-一个限制因素。先发制人的措施，以防止放射治疗相关的并发症，因此，成为常态。正在探索的清除自由基的氧，促进细胞再增殖，或增强DNA修复放射保护剂，以避免唾液功能减退8-11。 外分泌腺唾液腺分泌物流入通过主排泄管道口。个的口腔内插管E对于造影剂注射排泄管道作为门诊手术常规做。利用类似的方法，唾液腺可以直接针对局部治疗12。除了降低的全身性副作用的风险，retroductal腺滴注增加的好处。围绕导管树唾液腺细胞的单层布置允许指定所有唾液腺上皮细胞，以及腺充当屏障纤维封装，以减少不必要的治疗性传播。从本质上讲，唾液腺是最适宜于腺体的苦难，如放射性唾液腺功能减退的有针对性的治疗。 （ - 2.5格雷/分/天，每周五天1.8），为期两周的用于癌症治疗常规辐射中的小剂量输送。因此，无线电防护治疗，显示在实验模型对旷日持久的辐射方案的疗效有较大的临床影响。康博分次辐射后mised唾液腺功能已被记录在小动物，但辐射源，剂量分数，和使用的协议是多种多样9,10,13。 该报告建立了retroductal交付使用病人相关的辐射源和剂量率大鼠颌下腺局部放射方法。

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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.

适应微创技术造影，大唾液腺的局部治疗是可行的。在大鼠颌下腺唾液腺Retroductal给药试图通过沃顿管道口腔内插管（ 图2）。沃顿的唾液腺导管上位于口底的舌下乳头打开，但这些孔不容易看到。插管的插入物，因此，通过温和探测完成。为了避免不良出血或穿孔管，同时将套管没有使用武力。在导管插管的光滑无电阻的通道是由管子的平缓来回运动证实。 ...

Watch this Scientific Journal Video about Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction at JoVE.com

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

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颌下腺灌输和本地化分割照射在涎腺功能减退症大鼠模型

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

retroductal-submandibular-gland-instillation-localized-fractionated

Universidad de Cartagena UDEC

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JoVE Journal

Medicine

11.3K 次观看.  Louisiana State University Health Sciences Center. 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.

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

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

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their head and neck regions for cancer treatment. Both groups of patients experience symptoms such as xerostomia (dry mouth), dysphagia 
(impaired chewing and swallowing), severe dental caries, altered taste, oro-pharyngeal infections (candidiasis), mucositis, pain and discomfort.
One innovative approach of regenerative medicine for the treatment of salivary gland hypo-function is speculated in RS Redman, E Mezey 
et al. 2009: stem cells can be directly deposited by cannulation into the gland as a potent method in reviving the functions of the impaired organ. Presumably, 
the migrated foreign stem cells will differentiate into glandular cells to function as part of the host salivary gland. Also, this cannulation technique is an 
expedient and effective delivery method for clinical gene transfer application.
Here we illustrate the steps involved in performing the cannulation procedure on the mouse submandibular salivary gland via the Wharton's duct 
(Fig 1). C3H mice (Charles River, Montreal, QC, Canada) are used for this experiment, which have been kept under clean conventional conditions at 
the McGill University animal resource center. All experiments have been approved by the University Animal Care Committee and were in accordance with the 
guidelines of the Canadian Council on Animal Care.
For this experiment, a trypan blue solution is infused into the gland through the opening of the Wharton's duct using a 
insulin syringe with a 29-gauge needle encased inside a polyethylene tube. Subsequently, the mouse is dissected to show that the infusions migrated 
into the gland successfully.

A protocol for the cannulation of the mouse submandibular salivary gland via the Wharton's duct is described. For this experiment, the trypan blue solution is used as a dyer to demonstrate how this technique effectively delivers infusions into the targeted gland, and to suggest the reliability of this new approach as a potential clinical drug/cell therapy for the regeneration of salivary glands.

沃顿商学院的导管插管通过小鼠颌下腺唾液腺

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

科研

医学

Isolation of Mouse Salivary Gland Stem Cells

Mature salivary glands of both human and mouse origin comprise a minimum of five cell types, each of which facilitates the production and excretion of saliva into the oral cavity. Serous and mucous acinar cells are the protein and mucous producing factories of the gland respectively, and represent the origin of saliva production. Once synthesised, the various enzymatic and other proteinaceous components of saliva are secreted through a series of ductal cells bearing epithelial-type morphology, until the eventual expulsion of the saliva through one major duct into the cavity of the mouth. The composition of saliva is also modified by the ductal cells during this process.
In the manifestation of diseases such as Sj&ouml;gren's syndrome, and in some clinical situations such as radiotherapy treatment for head and neck cancers, saliva production by the glands is dramatically reduced 1,2. The resulting xerostomia, a subjective feeling of dry mouth, affects not only the ability of the patient to swallow and speak, but also encourages the development of dental caries and can be socially debilitating for the sufferer.
The restoration of saliva production in the above-mentioned clinical conditions therefore represents an unmet clinical need, and as such several studies have demonstrated the regenerative capacity of the salivary glands 3-5. Further to the isolation of stem cell-like populations of cells from various tissues within the mouse and human bodies 6-8, we have shown using the described method that stem cells isolated from mouse salivary glands can be used to rescue saliva production in irradiated salivary glands 9,10. This discovery paves the way for the development of stem cell-based therapies for the treatment of xerostomic conditions in humans, and also for the exploration of the salivary gland as a microenvironment containing cells with multipotent self-renewing capabilities.

An optimized protocol for the isolation of stem cells from the mouse salivary gland is described. The method employs enzymatic and mechanical digestion, and permits isolation of salispheres containing cells with characteristics of stem cells.

鼠标涎腺干细胞的分离

Mature salivary glands of both human and mouse origin comprise a minimum of five cell types, each of which facilitates the production and excretion of ...

生物学

Retroductal Nanoparticle Injection to the Murine Submandibular Gland

Two common goals of salivary gland therapeutics are prevention and cure of tissue dysfunction following either autoimmune or radiation injury. By locally delivering bioactive compounds to the salivary glands, greater tissue concentrations can be safely achieved versus systemic administration. Furthermore, off target tissue effects from extra-glandular accumulation of material can be dramatically reduced. In this regard, retroductal injection is a widely used method for investigating both salivary gland biology and pathophysiology. Retroductal administration of growth factors, primary cells, adenoviral vectors, and small molecule drugs has been shown to support gland function in the setting of injury. We have previously shown the efficacy of a retroductally injected nanoparticle-siRNA strategy to maintain gland function following irradiation. Here, a highly effective and reproducible method to administer nanomaterials to the murine submandibular gland through Wharton's duct is detailed (Figure 1). We describe accessing the oral cavity and outline the steps necessary to cannulate Wharton's duct, with further observations serving as quality checks throughout the procedure.

Local drug delivery to the submandibular glands is of interest in understanding salivary gland biology and for the development of novel therapeutics. We present an updated and detailed retroductal injection protocol, designed to improve delivery accuracy and experimental reproducibility. The application presented herein is the delivery of polymeric nanoparticles.

Retroductal 纳米微粒注射液对小鼠颌下腺的治疗

Two common goals of salivary gland therapeutics are prevention and cure of tissue dysfunction following either autoimmune or radiation injury. By locally ...

生物工程

A Method for the Measurement of Salivary Gland Function in Mice

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced saliva production. Similarly, saliva production is severely compromised in patients receiving radiation treatment for head and neck cancers. Rodent models, developed to mimic these clinical conditions, facilitate an understanding of the disease pathogenesis and allow for the development of new therapeutic strategies. Therefore, the ability to accurately, reproducibly, and repeatedly measure salivary gland function in animal models is critical. Building on procedures previously described in the literature, a method was developed that meets these criteria and was used to evaluate salivary gland function in mice. An additional advantage of this new method is that it is easily mastered, and has little inter-operator variation. Salivary gland function is evaluated as the amount (weight or volume) or rate (mL/min) of saliva produced in response to pilocarpine stimulation. The collected saliva is a good source for the analyses of protein content, immunoglobulin concentrations, and other biomolecules.

Salivary gland hypofunction is a frequent consequence of autoimmune disease and radiation therapy. Reproducible evaluation of salivary gland function in mouse models of these diseases is a technical challenge. Here, a simple method for accurate and reproducible measurement of saliva production in mice is described.

小鼠涎腺功能的测定方法

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced ...

免疫与感染

Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, including cell biology, oncology, dentistry, and immunology. The SMG is an exocrine gland comprised of secretory epithelial cells, myofibroblasts, endothelial cells, nerves, and extracellular matrix. Dynamic cellular processes in the rat and mouse SMG have previously been imaged, mostly using inverted multi-photon microscope systems. Here, we describe a straightforward protocol for the surgical preparation and stabilization of the murine SMG in anesthetized mice for in vivo imaging with upright multi-photon microscope systems. We present representative intravital image sets of endogenous and adoptively transferred fluorescent cells, including the labeling of blood vessels or salivary ducts and second harmonic generation to visualize fibrillar collagen. In sum, our protocol allows for surgical preparation of mouse salivary glands in upright microscopy systems, which are commonly used for intravital imaging in the field of immunology.

We describe a protocol to surgically expose and stabilize the murine submandibular salivary gland for intravital imaging using upright intravital microscopy. This protocol is easily adaptable to other exocrine glands of the head and neck region of mice and other small rodents.

直立活体显微术制备小鼠颌下腺涎腺

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, ...

Ultrasound-guided Botulinum Toxin-A Injections: A Method of Treating Sialorrhea

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an impaired quality of life, often worsened by correlated complications such as aspiration pneumonia, oral infections, dental caries, and maceration of the skin. Diverse therapeutic approaches have been proposed for the treatment of sialorrhea, including surgery and the use of anticholinergic agents, with limited results and the possible occurrence of serious adverse events. Recently, botulinum toxin (BoNT) injection within the major salivary glands has been proposed in patients refractory to anticholinergic therapy, with the aim of inhibiting local acetylcholine release and gland activity. 
In order to obtain a better outcome in terms of reduction of saliva production, efficacy, duration, and avoidance of major adverse events, we developed an ultrasound-guided BoNT-type A injection technique accurately described in the text. Here we present a method of treating sialorrhea with bilateral parotid and submandibular gland BoNT-type A injections under ultrasound guidance. Four quadrants of the parotid gland and two quadrants of the submandibular gland are visualized and injected using two accesses and one access, respectively. 
The ultrasound-guided procedure provides a simple, non-invasive, real-time visualization of the muscular and glandular tissues and their surrounding structures, optimizing treatment efficacy and safety.

Botulinum toxin is injected into the salivary glands for the treatment of sialorrhea. The ultrasound-guided botulinum toxin injection in both the parotid and submandibular glands demonstrates a sustained efficacy and the absence of serious side effects.

超声引导下肉毒毒素-A注射液:治疗流涎的一种方法

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an ...

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.

小鼠涎腺照射后通过匹属刺激进行唾液功能评估

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

Retrograde Parotid Gland Infusion through Stensen's Duct in a Non-Human Primate for Vectored Gene Delivery

Salivary glands are an attractive tissue target for gene therapy with promising results already leading to human trials. They are inherently capable of secreting proteins into the bloodstream and are easily accessible, making them potentially superior tissue sites for replacement hormone production or vaccination by gene transfer. Suggested methods for gene delivery include transcutaneous injection and retrograde infusion through salivary ducts. We demonstrate how to perform Retrograde Salivary Gland Infusion (RSGI) in non-human primates. We describe the important anatomic landmarks including identification of the parotid papilla, an atraumatic method of cannulating and sealing Stensen&#39;s Duct utilizing basic dental tools, polyethylene tubing, and cyanoacrylate, and the appropriate rate of infusion. While this is the least traumatic method of delivery, the method is still limited by the volume able to be delivered (&#60;0.5 mL) and the potential for trauma to the duct and gland. We demonstrate using fluoroscopy that an infusate can be fully delivered into the gland, and further demonstrate by immunohistochemistry the transduction of a typical vector and expression of the delivered gene.

Salivary glands have been proposed as a tissue target site for gene therapy, especially in the area of vaccination by gene transfer. We demonstrate gene delivery in a non-human primate model utilizing retrograde parotid infusion.

逆行帕罗蒂德格兰德输液通过斯滕森的管道在非人类灵长类病原体矢量基因交付

Salivary glands are an attractive tissue target for gene therapy with promising results already leading to human trials. They are inherently capable of ...

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.

下颌骨和腮腺表型模型对组织型培养的放射治疗是机体活力特征的关键

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

小鼠下颌下腺再生的实时实时成像

Interrogating Cell-Cell Interactions in the Salivary Gland via Ex Vivo Live Cell Imaging

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal role played by macrophages in driving the regenerative response. However, our understanding of this critical role has primarily relied on static views obtained from fixed tissue biopsies. To overcome this limitation and gain insights into these interactions in real time, this study outlines a comprehensive protocol for culturing salivary gland tissue ex vivo and capturing live images of cell migration.
The protocol involves several key steps: First, mouse submandibular salivary gland tissue is carefully sliced using a vibratome and then cultured at an air-liquid interface. These slices can be intentionally injured, for instance, through exposure to radiation, to induce cellular damage and trigger the regenerative response. To track specific cells of interest, they can be endogenously labeled, such as by utilizing salivary gland tissue collected from genetically modified mice where a particular protein is marked with green fluorescent protein (GFP). Alternatively, fluorescently-conjugated antibodies can be employed to stain cells expressing specific cell surface markers of interest. Once prepared, the salivary gland slices are subjected to live imaging using a high-content confocal imaging system over a duration of 12 h, with images captured at 15 min intervals. The resulting images are then compiled to create a movie, which can subsequently be analyzed to extract valuable cell behavior parameters. This innovative method provides researchers with a powerful tool to investigate and better understand macrophage interactions within the salivary gland following injury, thereby advancing our knowledge of the regenerative processes at play in this dynamic biological context.

作者聚焦:研究损伤后唾液腺再生中巨噬细胞-上皮细胞的相互作用 

Immunofluorescent imaging is constrained by the ability to observe complex, time-dependent biological processes in just a single snapshot in time. This study outlines a live-imaging approach conducted on precision-cut mouse submandibular gland slices. This approach allows for the real-time observation of cell-cell interactions during homeostasis and the processes of regeneration and repair.

通过离体活细胞成像询问唾液腺中的细胞间相互作用

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal ...