وأيد هذا العمل من المنح المقدمة من جمعية الألمانية للبحوث (KA1270/3-1/2) وBONFOR (O-112.0040).

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+inducible+nitric+oxide+synthase+in+postoperative+intestinal+smooth+muscle+dysfunction+in+rodents.">Role of inducible nitric oxide synthase in postoperative intestinal smooth muscle dysfunction in rodents.</a> Gastroenterology. 118, 316-327 (2000).</li><li>Iyer, S., Saunders, W. B., Stemkowski, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Economic+burden+of+postoperative+ileus+associated+with+colectomy+in+the+United+States.">Economic burden of postoperative ileus associated with colectomy in the United States.</a> J. Manag. Care Pharm. 15, 485-494 (2009).</li><li>Miller, M. S., Galligan, J. J., Burks, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+measurement+of+intestinal+transit+in+the+rat.">Accurate measurement of intestinal transit in the rat.</a> J. Pharmacol. Methods. 6, 211-217 (1981).</li><li>Kalff, J. C., Schraut, W. H., Simmons, R. L., Bauer, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+manipulation+of+the+gut+elicits+an+intestinal+muscularis+inflammatory+response+resulting+in+postsurgical+ileus.">Surgical manipulation of the gut elicits an intestinal muscularis inflammatory response resulting in postsurgical ileus.</a> Ann. Surg. 228, 652-663 (1998).</li><li>Wehner, S., Straesser, S., Vilz, T. O., Pantelis, D., Sielecki, T., de lC, V., Hirner, A., Kalff, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+p38+mitogen-activated+protein+kinase+pathway+as+prophylaxis+of+postoperative+ileus+in+mice.">Inhibition of p38 mitogen-activated protein kinase pathway as prophylaxis of postoperative ileus in mice.</a> Gastroenterology. 136, 619-629 (2009).</li><li>Konigsrainer, I., Turck, M. H., Eisner, F., Meile, T., Hoffmann, J., Kuper, M., Zieker, D., Glatzle, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Gut+is+not+only+the+Target+but+a+Source+of+Inflammatory+Mediators+Inhibiting+Gastrointestinal+Motility+During+Sepsis.">The Gut is not only the Target but a Source of Inflammatory Mediators Inhibiting Gastrointestinal Motility During Sepsis.</a> Cell Physiol. Biochem. 28, 753-760 (2011).</li><li>Schwarz, N. T., Kalff, J. C., Turler, A., Speidel, N., Grandis, J. R., Billiar, T. R., Bauer, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+jejunal+manipulation+causes+postoperative+pan-enteric+inflammation+and+dysmotility.">Selective jejunal manipulation causes postoperative pan-enteric inflammation and dysmotility.</a> Gastroenterology. 26, 159-169 (2004).</li><li>Engel, D. R., Koscielny, A., Wehner, S., Maurer, J., Schiwon, M., Franken, L., Schumak, B., Limmer, A., Sparwasser, T., Hirner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T+helper+type+1+memory+cells+disseminate+postoperative+ileus+over+the+entire+intestinal+tract.">T helper type 1 memory cells disseminate postoperative ileus over the entire intestinal tract.</a> Nat. Med. , (2010).</li><li>Stoffels, B., Schmidt, J., Nakao, A., Nazir, A., Chanthaphavong, R. S., Bauer, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+interleukin+10+in+murine+postoperative+ileus.">Role of interleukin 10 in murine postoperative ileus.</a> Gut. 58, 648-660 (2009).</li></ol>

الإعلان عن أي تضارب في المصالح.

كشفت دراسة مقارنة وتحليل مستويات مختلفة من IM (صغير اندحاق الامعاء لمدة 10 دقيقة مقابل بلطف التفتيش في الأمعاء الدقيقة باستخدام القطن تطبيقها مقابل اثنين IM مع إخلاء محتوى الأمعاء الدقيقة في الأعور) وجود علاقة بين مدى التلاعب الجراحية وPOI. مقارنة مع المجموعات الأخرى IM أ...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف / معدات </td><td> شركة </td><td> كتالوج رقم </td></tr><tr><td> Ethilon 5/0 </td><td> Ethicon </td><td> 1666H </td></tr><tr><td> القطن تطبيقها </td><td> شوهوا </td><td> 71010 </td></tr><tr><td> قسطرة الشرايين </td><td> Vygon </td><td> 115-798 </td></tr><tr><td> 96 لوحة جيدا (أسود) </td><td> غرينر بيو واحد </td><td> 655096 </td></tr><tr><td> الزجاج الكرة (2 مم) </td><td> Worf </td><td> متوفرة عند الطلب </td></tr><tr><td> دبابيس الحشرات </td><td> أدوات العلوم الجميلة </td><td> 26000-25 </td></tr><tr><td> تاق ييتس الكاشف </td><td> Polyscience </td><td> 560694 </td></tr><tr><td> Aquatek </td><td> ميرك </td><td> HC109850 </td></tr><tr><td> DMEM </td><td> Lonza </td><td> BE12 F-604 </td></tr><tr><td> FITC-ديكستران (MW 70000) </td><td> سيغما الدريتش </td><td> 46945-500MG-F </td></tr></tbody></table>

functional assessment of intestinal motility and gut wall inflammation in rodents: analyses in a standardized model of intestinal manipulation

التهاب القناة الهضمية هو السبب الشائع لمجموعة متنوعة من الأمراض التي تصيب الإنسان. نماذج البحوث الحيوانية حاسمة في التحقيق في مجمع الخلوية والجزيئية علم الأمراض المعوية. على الرغم من أن الغشاء المخاطي الغلالة وغالبا ما يكون الجهاز من الاهتمام في العديد من الأمراض الالتهابية، والأعمال الأخيرة أثبتت أن الظاهرة العضلية (ME) هو أيضا جهازا مناعيا للغاية التي تؤوي شبكة كثيفة من immunocytes المقيمين. 1،2 أجريت هذه الأعمال ضمن موحدة نموذج للتلاعب المعوية (IM) الذي يؤدي إلى التهاب في جدار الأمعاء، وتقتصر أساسا إلى ME. سريريا هذا الالتهاب يؤدي إلى الأمعاء لفترة طويلة dysmotility، والمعروفة باسم علوص ما بعد الجراحة (POI) والذي هو اختلاط متكررة والتي لا يمكن تجنبها بعد عملية جراحية في البطن. 3 ويتميز التهاب تحرير سطاء proinflammatory مثل IL-6 4 أو الناقلات العصبية IL-1β أو المثبطة مثل nitriج أكسيد (NO) .. 5 وفي وقت لاحق، وأرقام هائلة من immunocytes يتسرب إلى ME، التي يسيطر عليها العدلات النوى (السوداء) وحيدات والحفاظ أخيرا POI .. 2 دائم لعدة أيام، وهذا يؤدي إلى شلل الأمعاء زيادة خطر الطموح، إزفاء البكتيرية والمضاعفات المعدية حتى تعفن الدم والفشل العضوي المتعدد ويسبب عبئا عالية الاقتصادية .. 6 في هذا المخطوط أن نبرهن على نموذج موحد للتقييم في IM والجسم الحي العبور المعوي (GIT) والعبور القولون. وعلاوة على ذلك نحن يبرهن على وجود طريقة لفصل لي من الغشاء المخاطي تليها الغلالة تحليل المناعية، وهو أمر حاسم للتمييز بين الاستجابات الالتهابية في كل من هذه المقصورات immunoactive غاية جدار الامعاء. جميع التحليلات بسهولة نقلها إلى أي نماذج بحثية أخرى، مما يؤثر على وظيفة الجهاز الهضمي.

Watch this Scientific Journal Video about Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation at JoVE.com

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

علوص ما بعد الجراحة (POI) هي مضاعفات عملية جراحية في البطن مما يؤدي إلى زيادة معدلات الاعتلال والبقاء في المستشفى لفترات طويلة. لأن استراتيجيات وقائية أو علاجية تعاني من نقص البحوث المكثفة ضرورية. أنشئت لذلك لدينا نموذج الفأر موحدة وقابلة للتحقيق في الفيزيولوجيا المرضية لPOI ودراسة الخيارات العلاجية المحتملة.

وظيفية تقييم الحركة المعوية والتهاب الأمعاء الجدار في القوارض: التحليلات في نموذج موحد من التلاعب المعوية

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

functional-assessment-intestinal-motility-gut-wall-inflammation

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JoVE Journal

Medicine

21.9K Views.  University of Bonn. علوص ما بعد الجراحة (POI) هي مضاعفات عملية جراحية في البطن مما يؤدي إلى زيادة معدلات الاعتلال والبقاء في المستشفى لفترات طويلة. لأن استراتيجيات وقائية أو علاجية تعاني من نقص البحوث المكثفة ضرورية. أنشئت لذلك لدينا نموذج الفأر موحدة وقابلة للتحقيق في الفيزيولوجيا ...

Video: Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions preclinically. However, valid assessment of pathological alterations often requires histological analysis, and when performed ex vivo, necessitates death of the animal. Therefore in conventional experimental settings, intra-individual follow-up examinations are rarely possible. Thus, development of murine endoscopy in live mice enables investigators for the first time to both directly visualize the gastrointestinal mucosa and also repeat the procedure to monitor for alterations. Numerous applications for in vivo murine endoscopy exist, including studying intestinal inflammation or wound healing, obtaining mucosal biopsies repeatedly, and to locally administer diagnostic or therapeutic agents using miniature injection catheters. Most recently, molecular imaging has extended diagnostic imaging modalities allowing specific detection of distinct target molecules using specific photoprobes. In conclusion, murine endoscopy has emerged as a novel cutting-edge technology for diagnostic experimental in vivo imaging and may significantly impact on preclinical research in various fields.

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Small animal imaging techniques allow serial diagnostic examinations and therapeutic interventions in vivo. Recently, the scope of applications has significantly widened and currently includes assessment of colonic tumor development, wound healing and monitoring of inflammation. This protocol illustrates these diverse potential applications of murine endoscopy.

الفئران التنظير ل<em&gt; في فيفو</em&gt; شفاء المتعدد الوسائط التصوير من التسرطن وتقييم المعوية والتهاب الجرح

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

نموذج الفئران من المعوية الإسكيمية-ضخه إصابة

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not routinely employed in animal models. Conventional experimental readouts often rely on post mortem and ex vivo analyses, which prevent intra-individual follow-up examinations and increase the number of study animals needed. Fluorescence-mediated tomography enables the non-invasive, repetitive, quantitative, three-dimensional assessment of fluorescent probes. It is highly sensitive and permits the use of molecular makers, which allows for the specific detection and characterization of distinct molecular targets. In particular, targeted probes represent an innovative tool for analyzing gene activation and protein expression in inflammation, autoimmune disease, infection, vascular disease, cell migration, tumorigenesis, etc. In this article, we provide step-by-step instructions on this sophisticated imaging technology for the in vivo detection and characterization of inflammation (i.e., F4/80-positive macrophage infiltration) in a widely used murine model of intestinal inflammation. This technique might also be used in other research areas, such as immune cell or stem cell tracking.

Target-specific probes represent an innovative tool for analyzing molecular mechanisms, such as protein expression in various types of disease (e.g., inflammation, infection, and tumorigenesis). In this study, we describe a quantitative three-dimensional tomographic assessment of intestinal macrophage infiltration in a murine model of colitis using F4/80-specific fluorescence-mediated tomography.

الأسفار بوساطة التصوير المقطعي للكشف والتحديد الكمي لالتهاب الأمعاء مورين المتصلة بلعم

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

Intestinal Stem Cell Isolation and Culture in a Porcine Model of Segmental Small Intestinal Ischemia

Intestinal ischemia remains a major cause of morbidity and mortality in human and veterinary patients. Many disease processes result in intestinal ischemia, when the blood supply and therefore oxygen is decreased to the intestine. This leads to intestinal barrier loss and damage to the underlying tissue. Intestinal stem cells reside at the base of the crypts of Lieberk&#252;hn and are responsible for intestinal renewal during homeostasis and following injury. Ex vivo cell culture techniques have allowed for the successful study of epithelial stem cell interactions by establishing culture conditions that support the growth of three-dimensional epithelial organ-like systems (termed "enteroids" and "colonoids" from the small and large intestine, respectively). These enteroids are composed of crypt and villus-like domains and mature to contain all of the cell types found within the epithelium. Historically, murine models have been utilized to study intestinal injury. However, a porcine model offers several advantages including similarity of size as well as gastrointestinal anatomy and physiology to that of humans. By utilizing a porcine model, we establish a protocol in which segmental loops of intestinal ischemia can be created within a single animal, enabling the study of differing time points of ischemic injury and repair in vivo. Additionally, we describe a method to isolate and culture the intestinal stem cells from the ischemic loops of intestine, allowing for the continued study of epithelial repair, modulated by stem cells, ex vivo.

This protocol will enable readers to successfully establish a porcine model of segmental intestinal ischemia and subsequently isolate and culture intestinal stem cells for the study of epithelial repair following injury.

عزل الخلايا الجذعية المعوية والثقافة في نموذج الخنزير من المجزأ الاسكيمية الأمعاء الصغيرة

Intestinal ischemia remains a major cause of morbidity and mortality in human and veterinary patients. Many disease processes result in intestinal ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

استخدام خزعة كيميائية لتقييم جودة الكسب غير المشروع

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...

Standardized Histomorphometric Evaluation of Osteoarthritis in a Surgical Mouse Model

One of the most prevalent joint disorders in the United States, osteoarthritis (OA) is characterized by progressive degeneration of articular cartilage, primarily in the hip and knee joints, which results in significant impacts on patient mobility and quality of life. To date, there are no existing curative therapies for OA able to slow down or inhibit cartilage degeneration. Presently, there is an extensive body of ongoing research to understand OA pathology and discover novel therapeutic approaches or agents that can efficiently slow down, stop, or even reverse OA. Thus, it is crucial to have a quantitative and reproducible approach to accurately evaluate OA-associated pathological changes in the joint cartilage, synovium, and subchondral bone. Currently, OA severity and progression are primarily assessed using the Osteoarthritis Research Society International (OARSI) or Mankin scoring systems. In spite of the importance of these scoring systems, they are semiquantitative and can be influenced by user subjectivity. More importantly, they fail to accurately evaluate subtle, yet important, changes in the cartilage during the early disease states or early treatment phases. The protocol we describe here uses a computerized and semiautomated histomorphometric software system to establish a standardized, rigorous, and reproducible quantitative methodology for the evaluation of joint changes in OA. This protocol presents a powerful addition to the existing systems and allows for more efficient detection of pathological changes in the joint.

The current protocol establishes a rigorous and reproducible method for quantification of morphological joint changes that accompany osteoarthritis. Application of this protocol can be valuable in monitoring disease progression and evaluating therapeutic interventions in osteoarthritis.

تقييم قياسي للفزة الهيستومترية لهشاشة العظام في نموذج الماوس الجراحي

One of the most prevalent joint disorders in the United States, osteoarthritis (OA) is characterized by progressive degeneration of articular cartilage, ...

A Murine Model of Fetal Exposure to Maternal Inflammation to Study the Effects of Acute Chorioamnionitis on Newborn Intestinal Development

Chorioamnionitis is a common precipitant of preterm birth and is associated with many of the morbidities of prematurity, including necrotizing enterocolitis (NEC). However, a mechanistic link between these two conditions remains yet to be discovered. We have adopted a murine model of chorioamnionitis involving lipopolysaccharide (LPS)-induced fetal exposure to maternal inflammation (FEMI). This model of FEMI induces a sterile maternal, placental, and fetal inflammatory cascade, which is also present in many cases of clinical chorioamnionitis. Although models exist that utilize live bacteria and more accurately mimic the pathophysiology of an ascending infection resulting in chorioamnionitis, these methods may cause indirect effects on development of the immature intestinal tract and the associated developing microbiome. Using this protocol, we have demonstrated that LPS-induced FEMI results in a dose-dependent increase in pregnancy loss and preterm birth, as well as disruption of normal intestinal development in offspring. Further, we have demonstrated that FEMI significantly increases intestinal injury and serum cytokines in offspring, while simultaneously decreasing goblet and Paneth cells, both of which provide a first line of innate immunity against intestinal inflammation. Although a similar model of LPS-induced FEMI has been used to model the association between chorioamnionitis and subsequent abnormalities of the central nervous system, to our knowledge, this protocol is the first to attempt to elucidate a mechanistic link between chorioamnionitis and later perturbations in intestinal development as a potential link between chorioamnionitis and NEC.

We developed a model of chorioamnionitis to simulate fetal exposure to maternal inflammation (FEMI) without complications of live organisms to examine the effects of FEMI on development of the offspring&#8217;s intestinal tract. This allows for study of mechanistic causes for development of intestinal injury following chorioamnionitis.

نموذج مورين للتعرض الجنين لالتهاب الأم لدراسة آثار التهاب المشيمة الحاد على نمو الأمعاء حديثي الولادة

Chorioamnionitis is a common precipitant of preterm birth and is associated with many of the morbidities of prematurity, including necrotizing ...

Induction of Intestinal Inflammation by Adoptive Transfer of CBir1 TCR Transgenic CD4+ T Cells to Immunodeficient Mice

With the increase of incidence, inflammatory bowel diseases (IBD), which are chronic diseases affecting the gastrointestinal tract, impose a considerable health and financial burden on individuals and society. Therefore, it is critical to investigate the mechanisms underlying the pathogenesis and development of IBD. Here, a gut microbiota antigen-specific T cell transfer colitis model is described. CBir1 flagellin has been recognized as the immunodominant gut bacterial antigen in experimental colitis and patients with Crohn&#39;s disease. CBir1 TCR transgenic na&#970;ve CD4+ T cells, specific to CBir1 flagellin, can induce chronic colitis after adoptive transfer into immune-deficient Rag1-/- mice. The disease severity is assessed by histopathology. The CD4+ T cell phenotypes in colonic lamina propria are also determined. This model closely resembles the development of IBD, which provides an ideal murine model for investigating the mechanisms driving the pathogenesis of IBD and testing the potential drugs for treating IBD.

Induction of Intestinal Inflammation by Adoptive Transfer of CBir1 TCR Transgenic CD4+ T Cells to Immunodeficient Mice

In this protocol, a gut microbiota antigen-specific T cell adoptive transfer colitis model is described. CD4+ T cells are isolated from CBir1 TCR transgenic mice. These are specific for an immunodominant gut microbiota antigen CBir1 flagellin, which is transferred into recipient Rag1-/- mice, leading to intestinal inflammation.

تحريض التهاب الأمعاء عن طريق النقل المتبني للخلايا التائية المعدلة وراثيا CBir1 TCR CD4 + إلى الفئران التي تعاني من نقص المناعة

With the increase of incidence, inflammatory bowel diseases (IBD), which are chronic diseases affecting the gastrointestinal tract, impose a considerable ...

Isolation of Murine Intestinal Mesenchyme Resulting in a High Yield of Telocytes

The murine small intestine, or colon mesenchyme, is highly heterogenous, containing distinct cell types including blood and lymphatic endothelium, nerves, fibroblasts, myofibroblasts, smooth muscle cells, immune cells, and the recently identified cell type, telocytes. Telocytes are unique mesenchymal cells with long cytoplasmic processes, reaching a distance of tens to hundreds of microns from the cell body. Telocytes have recently emerged as an important intestinal stem cell niche component, providing Wnt proteins that are essential for stem and progenitor cell proliferation.
Although protocols on how to isolate mesenchyme from the mouse intestine are available, it is not clear whether these procedures allow the efficient isolation of telocytes. Isolating telocytes efficiently requires special protocol adjustments that would allow dissociation of the strong cell-cell contact between telocytes and neighboring cells without affecting their viability. Here, available intestinal mesenchyme isolation protocols were adjusted to support the successful isolation and culture of mesenchyme containing a relatively high yield of viable single-cell telocytes.
The obtained single-cell suspension can be analyzed by several techniques, such as immunostaining, cell sorting, imaging, and mRNA experiments. This protocol yields mesenchyme with sufficiently conserved antigenic and functional properties of telocytes, and can be used for several applications. For example, they can be used for co-culture with mouse- or human-derived organoids to support organoid growth with no growth factor supplementation, to better reflect the situation in the original tissue.

Here, we present a protocol to isolate murine intestinal mesenchyme, including telocytes. These can be used for several applications, such as co-culture with mouse or human-derived organoids, to support growth and better reflect the situation in the original tissue.

عزل اللحمة المتوسطة المعوية للفئران مما يؤدي إلى غلة عالية من الخلايا التيلوسيتية

The murine small intestine, or colon mesenchyme, is highly heterogenous, containing distinct cell types including blood and lymphatic endothelium, nerves, ...

التلاعب Tuina موحدة لهشاشة العظام في الركبة في الفئران

Tuina Manipulation to Reduce Inflammation and Cartilage Loss in Knee Osteoarthritis Rats

Clinical trials suggest that Tuina manipulation is effective in treating knee osteoarthritis (KOA), while further studies are required to discover its mechanism. Therefore, the manipulation of animal models of knee osteoarthritis is critical. This protocol provides a standard process for Tuina manipulation on KOA rats and a preliminary exploration of the mechanism of Tuina for KOA. The press and kneading manipulation method (a kind of Tuina manipulation that refers to pressing and kneading the specific area of the body surface) is applied on 5 acupoints around the knee joint of rats. The force and frequency of the manipulation were standardized by finger pressure recordings, and the position of the rat during manipulation is described in detail in the protocol. The effect of manipulation can be measured by pain behavior tests and microscopic findings in synovial and cartilage. KOA rats showed significant improvement in pain behavior. The synovial tissue inflammatory infiltration was reduced in the Tuina group, and the expression of tumor necrosis factor (TNF)-&#945; was significantly lower. Compared to the control group, chondrocyte apoptosis was less in the Tuina group. This study provides a standardized protocol for Tuina manipulation on KOA rats and preliminary proof that the therapeutic effects of Tuina may be related to reducing synovial inflammation and delayed chondrocyte apoptosis.

تسليط الضوء على المؤلف: فهم آلية التلاعب Tuina في الفئران 

We present a protocol for Tuina manipulation performed on plaster-immobilized-induced knee osteoarthritis rats. Based on the preliminary results, it suggested that the efficacy of the method relied on the reduction of inflammation and cartilage loss.

التلاعب Tuina للحد من الالتهاب وفقدان الغضروف في الفئران هشاشة العظام في الركبة

Clinical trials suggest that Tuina manipulation is effective in treating knee osteoarthritis (KOA), while further studies are required to discover its ...