המחברים מבקשים להודות Chiyoko קוביאשי ו Kiyokazu אגתה עבור עצות מועילות על השתלת planarian כמו גם חברים בעבר ובהווה של המעבדה סאנצ לדיונים יסולא בפז במהלך הפיתוח של טכניקות אלה. עבודה זו נתמכה על ידי NIH הדרכה גרנט (5T32 HD0791) כדי OCG ו NIH R37GM057260 את אס&quot;א. אס&quot;א הוא הווארד יוז חוקר רפואי במכון.

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Zo&#246;l. 3, 269-294 (1906).</li><li>Dubois, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+&#225;+l+'&#232;tude+de+la+migration+des+cellules+de+règènèration+chez+les+Planaires+dulcicoles.">Contribution &#225; l '&#232;tude de la migration des cellules de règènèration chez les Planaires dulcicoles.</a> Bull. Biol. Fr. Belg. 83, 213-283 (1949).</li><li>Purton, L. E., Scadden, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limiting+factors+in+murine+hematopoietic+stem+cell+assays.">Limiting factors in murine hematopoietic stem cell assays.</a> Cell Stem Cell. 1, 263-270 (2007).</li><li>Cebria, F., Newmark, P. 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Dyn. 238, 443-450 (2009).</li><li>Hayashi, T., Asami, M., Higuchi, S., Shibata, N., Agata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+planarian+X-ray-sensitive+stem+cells+by+fluorescence-activated+cell+sorting.">Isolation of planarian X-ray-sensitive stem cells by fluorescence-activated cell sorting.</a> Dev. Growth Differ. 48, 371-380 (2006).</li><li>Reddien, P. W., Oviedo, N. J., Jennings, J. R., Jenkin, J. 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אין ניגוד עניינים הצהיר.

החשיבות של חוסר תנועה Immobilization הוא ללא ספק השלב הקריטי ביותר על ההשלמה המוצלחת של כל התהליכים האלה. אם planaria הם משותקים באופן לא תקין לפני הקרנה חלקית, הם עלולים לעבור מתחת למגן את ההובלה, ייצור תוצאות לא עקביות ולא בלבול. בנוסף, אם בע?...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> שם מגיב </td><td> חברה </td><td> מספר קטלוגי </td></tr><tr><td> כללי העברת מטרת פיפטה </td><td> Samco </td><td> 691 </td></tr><tr><td> צינורות נימי (0.75 מ&quot;מ ID) </td><td> Fhc </td><td> 30-30-0 </td></tr><tr><td> צינורות נימי (OD 0.7 מ&quot;מ) </td><td> Fhc </td><td> 30-50-08 </td></tr><tr><td> Parafilm M </td><td> VWR </td><td> 52858-076 </td></tr><tr><td> Kimwipes 34155 </td><td> VWR </td><td> 500029-891 </td></tr><tr><td> מסנן נייר שחור </td><td> שלייכר &amp; Schuell </td><td> 10310809 </td></tr><tr><td> Whatman # 2 מסנן נייר 1002-055 </td><td> פישר סיינטיפיק </td><td> 09-810B </td></tr><tr><td> Whatman # 3 מסנן נייר 1003-185 </td><td> פישר סיינטיפיק </td><td> 09-820E </td></tr><tr><td> סיגריה מתגלגל נייר </td><td> זיג זג, מקורי </td><td> NA </td></tr><tr><td> פטרימנות </td><td> VWR </td><td> 82050-544 </td></tr><tr><td> מלקחיים דומונט, Inox # 5 </td><td> FST </td><td> 11251-20 </td></tr><tr><td> Chloretone </td><td> סיגמא אולדריץ&#39; </td><td> 112054 </td></tr><tr><td> קזאין </td><td> סיגמא אולדריץ&#39; </td><td> C3400 </td></tr><tr><td> להוביל שילדס </td><td> אלפא מערכות קורפ, Bluffdale, UT </td><td> עיצוב מותאם אישית </td></tr><tr><td> XRAD-320 ביולוגית Irradiator </td><td> Precision X-Ray, צפון Branford, CT </td><td> NA </td></tr></tbody></table>

planarian immobilization, partial irradiation, and tissue transplantation

Planarian, השטוחה מים מתוקים, הוכיחה להיות מערכת רב עוצמה לנתח התחדשות מטזואניים ו תא גזע 1,2 ביולוגיה. התחדשות Planarian של כל רקמות חסרים או פגומים מתאפשרת על ידי בתאי גזע בוגרים כינה neoblasts 3. למרות אלה בתאי גזע הוכחו בוודאות להיות pluripotent ומסוגלים לשחזר להפליא של כל בעל חיים 4, ההטרוגניות בקרב האוכלוסייה לתאי גזע ואת הדינמיקה של התנהגות הסלולר שלהם נותרו לא פתורות, במידה רבה. בשל ריבוי והפצה רחב של בתאי גזע ברחבי תוכנית הגוף planarian, שיטות מתקדמות של subpopulations מניפולציה בתאי גזע לצורך מחקר מולקולרי ומתפקד in vivo יש צורך. השתלת רקמות הקרנה חלקית שתי שיטות שבו subpopulation בתאי גזע planarian ניתן לבודד למחקר נוסף. הטכניקה לכל אחד יש יתרונות ברורים. Tהשתלת הנושא מאפשר הקדמה של בתאי גזע, לתוך שורה תמימה, כי הם או מטבעו שונה גנטית או טופלו בעבר פרמקולוגית. לחלופין, הקרנה חלקית מאפשר בידוד בתאי גזע בתוך הפונדקאי, זה לצד זה לרקמות ללא בתאי גזע, ללא הקדמה של פצע או עכוז של שלמות רקמות. באמצעות שתי השיטות, ניתן לחקור את הגורמים תאים אוטונומיים ולא אוטונומיים הנובעות בקרת התא פונקציות, כגון התפשטות, בידול, והגירה. הן רקמות להשתלה 5,6 ו חלקית קרינה 7 שימשו היסטורית בהגדרת רבים של שאלות על התחדשות planarian שנותרו תחת מחקר היום. עם זאת, טכניקות אלה נשארו underused בשל אופי מייגע ובלתי עקבית של השיטות הקודמות. הפרוטוקולים המוצגים כאן מייצגים צעד גדול קדימה בהפחתת זמןND המאמץ הנדרש כדי reproducibly לייצר מספר רב של בעלי חיים מורכבים או מוקרן באופן חלקי עם efficacies מתקרב 100 אחוז. אנו מכסים את כל התרבות של בעלי חיים גדולים, חוסר תנועה, לקראת הקרנת חלקית, השתלת רקמות, אופטימיזציה של התאוששות בעלי חיים. יתר על כן, העבודה המתוארת כאן ממחיש את היישום הראשון של שיטת הקרנה חלקית לשימוש עם planarian ביותר למד נרחב, Schmidtea mediterranea. בנוסף, רקמות יעיל השתלת ב planaria פותחת את הדלת לבדיקה תפקודית של subpopulations של בתאי גזע נאיבי או מטופלים מבחני repopulation, אשר כבר זמן רב את שיטת תקן הזהב של מנסה לאמוד את הפוטנציאל למבוגרים בתאי גזע אצל יונקים 8. אימוץ רחב של שיטות אלו יהיה ללא ספק להוביל להבנה טובה יותר של התנהגויות הסלולר של בתאי גזע בוגרים במהלך הומאוסטזיס רקמה והתחדשות.

Watch this Scientific Journal Video about Planarian Immobilization, Partial Irradiation, and Tissue Transplantation at JoVE.com

Planarian Immobilization, Partial Irradiation, and Tissue Transplantation

השיטה יעילה השתלת רקמות בגודל מוגדרת ועקבית בין planaria מתואר. היא כללה גם תיאור של איך להשתמש בטכניקה immobilization להשתלה ניתן להתאים, בשיתוף עם מגן עופרת, הקרנה חלקית של בעלי חיים.

Immobilization Planarian, הקרנה חלקית, השתלת רקמות

The planarian, a freshwater flatworm, has proven to be a powerful system for dissecting metazoan regeneration and stem cell biology1,2. Planarian ...

planarian-immobilization-partial-irradiation-tissue

Research

JoVE Journal

Biology

19.5K Views.  University of Utah School of Medicine. השיטה יעילה השתלת רקמות בגודל מוגדרת ועקבית בין planaria מתואר. היא כללה גם תיאור של איך להשתמש בטכניקה immobilization להשתלה ניתן להתאים, בשיתוף עם מגן עופרת, הקרנה חלקית של בעלי חיים.

Video: Planarian Immobilization, Partial Irradiation, and Tissue Transplantation

Orthotopic Hind-Limb Transplantation in Rats

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel therapeutic entity in reconstructive surgery. However, long term high-dose multi-drug immunosuppressive therapy is required to ensure graft survival, bearing the risk of serious side effects which halters broader application. Further progression in this field may depend on better understanding of basic immunology and ischemia reperfusion injury in composite tissue grafts.
To date, orthotopic hind limb transplantation in rats has been the preferred rodent model for reconstructive transplantation (RT), however, it is an extremely demanding procedure that requires extraordinary microsurgical skills for reattachment of vasculature, bones, muscles and nerves.
We have introduced the vascular cuff anastomosis technique to this model, providing a rapid and reliable approach to rat hind limb transplantation. This technique simplifies and shortens the surgical procedure and enables surgeons with basic microsurgical experience to successfully perform the operation with high survival and low complication rates. The technique seems to be well suited for immunological as well as ischemia reperfusion injury (IRI) studies.

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

השתלת orthotopic לימב הינד-בחולדות

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel ...

Immunology and Infection

Use of Human Perivascular Stem Cells for Bone Regeneration

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering1-3. PSCs are a FACS-sorted population of 'pericytes' (CD146+CD34-CD45-) and 'adventitial cells' (CD146-CD34+CD45-), each of which we have previously reported to have properties of mesenchymal stem cells. PSCs, like MSCs, are able to undergo osteogenic differentiation, as well as secrete pro-osteogenic cytokines1,2. In the present protocol, we demonstrate the osteogenicity of PSCs in several animal models including a muscle pouch implantation in SCID (severe combined immunodeficient) mice, a SCID mouse calvarial defect and a femoral segmental defect (FSD) in athymic rats. The thigh muscle pouch model is used to assess ectopic bone formation. Calvarial defects are centered on the parietal bone and are standardly 4 mm in diameter (critically sized)8. FSDs are bicortical and are stabilized with a polyethylene bar and K-wires4. The FSD described is also a critical size defect, which does not significantly heal on its own4. In contrast, if stem cells or growth factors are added to the defect site, significant bone regeneration can be appreciated. The overall goal of PSC xenografting is to demonstrate the osteogenic capability of this cell type in both ectopic and orthotopic bone regeneration models.

Human perivascular stem cells (PSCs) are a novel stem cell class for skeletal tissue regeneration similar to mesenchymal stem cells (MSCs). PSCs can be isolated by FACS (fluorescence activated cell sorting) from adipose tissue procured during standard liposuction procedures, then combined with an osteoinductive scaffold to achieve bone formation in vivo.

שימוש בתאי גזע לשחזור עצם Perivascular

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering1-3. PSCs are ...

Bioengineering

Transplantation of Zebrafish Pediatric Brain Tumors into Immune-competent Hosts for Long-term Study of Tumor Cell Behavior and Drug Response

Tumor cell transplantation is an important technique to define the mechanisms controlling cancer cell growth, migration, and host response, as well as to assess potential patient response to therapy. Current methods largely depend on using syngeneic or immune-compromised animals to avoid rejection of the tumor graft. Such methods require the use of specific genetic strains that often prevent the analysis of immune-tumor cell interactions and/or are limited to specific genetic backgrounds. An alternative method in zebrafish takes advantage of an incompletely developed immune system in the embryonic brain before 3 days, where tumor cells are transplanted for use in short-term assays (i.e., 3 to 10 days). However, these methods cause host lethality, which prevents the long-term study of tumor cell behavior and drug response. This protocol describes a simple and efficient method for the long-term orthotopic transplantation of zebrafish brain tumor tissue into the fourth ventricle of a 2-day-old immune-competent zebrafish. This method allows: 1) long-term study of tumor cell behaviors, such as invasion and dissemination; 2) durable tumor response to drugs; and 3) re-transplantation of tumors for the study of tumor evolution and/or the impact of different host genetic backgrounds. In summary, this technique allows cancer researchers to assess engraftment, invasion, and growth at distant sites, as well as to perform chemical screens and cell competition assays over many months. This protocol can be extended to studies of other tumor types and can be used to elucidate mechanisms of chemoresistance and metastasis.

The transplantation of cancer cells is an important tool for the identification of cancer mechanisms and therapeutic responses. Current techniques depend on immune-incompetent animals. Here, we describe a method to transplant zebrafish tumor cells into immune-competent embryos for the long-term analysis of tumor cell behavior and in vivo drug responses.

השתלת דג הזברה גידולים מוחיים במוח לתוך מארחים המוסמכת החיסונית עבור מחקר לטווח ארוך של התנהגות תאים סרטניים והתגובה לסמים

Tumor cell transplantation is an important technique to define the mechanisms controlling cancer cell growth, migration, and host response, as well as to ...

Cancer Research

Surgical Ablation Assay for Studying Eye Regeneration in Planarians

In the study of adult stem cells and regenerative mechanisms, planarian flatworms are a staple in vivo model system. This is due in large part to their abundant pluripotent stem cell population and ability to regenerate all cell and tissue types after injuries that would be catastrophic for most animals. Recently, planarians have gained popularity as a model for eye regeneration. Their ability to regenerate the entire eye (comprised of two tissue types: pigment cells and photoreceptors) allows for the dissection of the mechanisms regulating visual system regeneration. Eye ablation has several advantages over other techniques (such as decapitation or hole punch) for examining eye-specific pathways and mechanisms, the most important of which is that regeneration is largely restricted to eye tissues alone. The purpose of this video article is to demonstrate how to reliably remove the planarian optic cup without disturbing the brain or surrounding tissues. The handling of worms and maintenance of an established colony is also described. This technique uses a 31 G, 5/16-inch insulin needle to surgically scoop out the optic cup of planarians immobilized on a cold plate. This method encompasses both single and double eye ablation, with eyes regenerating within 1-2 weeks, allowing for a wide range of applications. In particular, this ablation technique can be easily combined with pharmacological and genetic (RNA interference) screens for a better understanding of regenerative mechanisms and their evolution, eye stem cells and their maintenance, and phototaxic behavioral responses and their neurological basis.

This protocol shows how to consistently excise planarian eyes (optic cups) without disturbing surrounding tissues. Using an insulin needle and syringe, either one or both eyes can be ablated to facilitate investigations into the mechanisms regulating eye regeneration, the evolution of visual regeneration, and the neural basis of light-induced behavior.

אבלציה כירורגית Assay עבור לימוד Regeneration Eye ב Planarians

In the study of adult stem cells and regenerative mechanisms, planarian flatworms are a staple in vivo model system. This is due in large part to their ...

Developmental Biology

Immobilization of Live Caenorhabditis elegans Individuals Using an Ultra-thin Polydimethylsiloxane Microfluidic Chip with Water Retention

Radiation is widely used for biological applications and for ion-beam breeding, and among these methods, microbeam irradiation represents a powerful means of identifying radiosensitive sites in living organisms. This paper describes a series of on-chip immobilization methods developed for the targeted microbeam irradiation of live individuals of Caenorhabditis elegans. Notably, the treatment of the polydimethylsiloxane (PDMS) microfluidic chips that we previously developed to immobilize C. elegans individuals without the need for anesthesia is explained in detail. This chip, referred to as a worm sheet, is resilient to allow the microfluidic channels to be expanded, and the elasticity allows animals to be enveloped gently. Also, owing to the self-adsorption capacity of the PDMS, animals can be sealed in the channels by covering the surface of the worm sheet with a thin cover film, in which animals are not pushed into the channels for enclosure. By turning the cover film over, we can easily collect the animals. Furthermore, the worm sheet shows water retention and allows C. elegans individuals to be subjected to microscopic observation for long periods under live conditions. In addition, the sheet is only 300 &#181;m thick, allowing heavy ions such as carbon ions to pass through the sheet enclosing the animals, thus allowing the ion particles to be detected and the applied radiation dose to be measured accurately. Because selection of the cover films used for enclosing the animals is very important for successful long-term immobilization, we conducted the selection of the suitable cover films and showed a recommended one among some films. As an application example of the chip, we introduced imaging observation of muscular activities of animals enclosing the microfluidic channel of the worm sheet, as well as the microbeam irradiation. These examples indicate that the worm sheets have greatly expanded the possibilities for biological experiments.

Immobilization of Live Caenorhabditis elegans Individuals Using an Ultra-thin Polydimethylsiloxane Microfluidic Chip with Water Retention

A series of immobilization methods has been established to allow the targeted irradiation of live Caenorhabditis elegans individuals using a recently developed ultra-thin polydimethylsiloxane microfluidic chip with water retention. This novel on-chip immobilization is also adequate for imaging observations. The detailed treatment and application examples of the chip are explained.

קיבעון של אנשים חיים Caenorhabditis elegans באמצעות שבב Microfluidic של דקים ועומדים בטמפרטורות Polydimethylsiloxane עם החזקת מים

Radiation is widely used for biological applications and for ion-beam breeding, and among these methods, microbeam irradiation represents a powerful means ...

Pharmacological and Functional Genetic Assays to Manipulate Regeneration of the Planarian Dugesia japonica

Free-living planarian flatworms have a long history of experimental usage owing to their remarkable regenerative abilities1. Small fragments excised from these animals reform the original body plan following regeneration of missing body structures. For example if a 'trunk' fragment is cut from an intact worm, a new 'head' will regenerate anteriorly and a 'tail' will regenerate posteriorly restoring the original 'head-to-tail' polarity of body structures prior to amputation (Figure 1A).
Regeneration is driven by planarian stem cells, known as 'neoblasts' which differentiate into ~30 different cell types during normal body homeostasis and enforced tissue regeneration. This regenerative process is robust and easy to demonstrate. Owing to the dedication of several pioneering labs, many tools and functional genetic methods have now been optimized for this model system. Consequently, considerable recent progress has been made in understanding and manipulating the molecular events underpinning planarian developmental plasticity2-9.
The planarian model system will be of interest to a broad range of scientists. For neuroscientists, the model affords the opportunity to study the regeneration of an entire nervous system, rather than simply the regrowth/repair of single nerve cell process that typically are the focus of study in many established models. Planarians express a plethora of neurotransmitters10, represent an important system for studying evolution of the central nervous system11, 12 and have behavioral screening potential13, 14. 
Regenerative outcomes are amenable to manipulation by pharmacological and genetic apparoaches. For example, drugs can be screened for effects on regeneration simply by placing body fragments in drug-containing solutions at different time points after amputation. The role of individual genes can be studied using knockdown methods (in vivo RNAi), which can be achieved either through cycles of microinjection or by feeding bacterially-expressed dsRNA constructs8, 9, 15. Both approaches can produce visually striking phenotypes at high penetrance- for example, regeneration of bipolar animals16-21. To facilitate adoption of this model and implementation of such methods, we showcase in this video article protocols for pharmacological and genetic assays (in vivo RNAi by feeding) using the planarian Dugesia japonica.

Pharmacological and Functional Genetic Assays to Manipulate Regeneration of the Planarian Dugesia japonica

An attractive model for studying stem cell differentiation within a live animal is the planarian flatworm. Regeneration is studied by simple amputation experiments that are easily performed in a basic laboratory and are amenable to pharmacological and genetic (in vivo RNAi) manipulation as detailed by protocols in this article.

מבחני גנטית תרופתי ו פונקציונלית לתפעל התחדשות של Planarian Japonica Dugesia</em

Free-living planarian flatworms have a long history of experimental usage owing to their remarkable regenerative abilities1. Small fragments excised from ...

Targeted and Selective Treatment of Pluripotent Stem Cell-derived Teratomas Using External Beam Radiation in a Small-animal Model

The growing number of victims of &#34;stem cell tourism,&#34; the unregulated transplantation of stem cells worldwide, has raised concerns about the safety of stem cell transplantation. Although the transplantation of differentiated rather than undifferentiated cells is common practice, teratomas can still arise from the presence of residual undifferentiated stem cells at the time of transplant or from spontaneous mutations in differentiated cells. Because stem cell therapies are often delivered into anatomically sensitive sites, even small tumors can be clinically devastating, resulting in blindness, paralysis, cognitive abnormalities, and cardiovascular dysfunction. Surgical access to these sites may also be limited, leaving patients with few therapeutic options. Controlling stem cell misbehavior is, therefore, critical for the clinical translation of stem cell therapy.
External beam radiation offers an effective means of delivering targeted therapy to decrease the teratoma burden while minimizing injury to surrounding organs. Additionally, this method avoids genetic manipulation or viral transduction of stem cells-which are associated with additional clinical safety and efficacy concerns. Here, we describe a protocol to create pluripotent stem cell-derived teratomas in mice and to apply external beam radiation therapy to selectively ablate these tumors in vivo.

Research on treatment strategies for pluripotent stem cell-derived teratomas is important for the clinical translation of stem cell therapy. Here, we describe a protocol to, first, generate stem cell-derived teratomas in mice and, then, to selectively target and treat these tumors in vivo using a small-animal irradiator.

טיפול ממוקד, סלקטיבי של Teratomas נגזר תאי גזע Pluripotent באמצעות קרינה חיצוני קרן במודל של חיית-קטן

The growing number of victims of &#34;stem cell tourism,&#34; the unregulated transplantation of stem cells worldwide, has raised concerns about the ...

Medicine

Partial Heterotopic Hindlimb Transplantation Model in Rats

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities after a complex tissue defect. Face and hand transplantations have changed disfigured patients' lives, giving them a new aesthetic and functional social organ. Despite promising outcomes, VCA is still underperformed due to life-long immunosuppression comorbidities and infectious complications. The rat is an ideal animal model for in vivo studies investigating immunological pathways and graft rejection mechanisms. Rats are also widely used in novel composite tissue graft preservation techniques, including perfusion and cryopreservation studies. Models used for VCA in rats must be reproducible, reliable, and efficient with low postoperative morbidity and mortality. Heterotopic limb transplantation procedures fulfill these criteria and are easier to perform than orthotopic limb transplants. Mastering rodent microsurgical models requires solid experience in microsurgery and animal care. Herein is reported a reliable and reproducible model of partial heterotopic osteomyocutaneous flap transplantation in rats, the postoperative outcomes, and the means of prevention of potential complications.

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

מודל השתלת הינדימבה הטרוטופית חלקית בחולדות

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities ...

Modified Heterotopic Hindlimb Osteomyocutaneous Flap Model in the Rat for Translational Vascularized Composite Allotransplantation Research

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand and face transplants and, more recently, abdominal wall, uterus, and urogenital transplants. Functional outcomes have exceeded initial expectations, and most recipients enjoy an improved quality of life. However, as clinical experience accumulates, chronic rejection and complications from the immunosuppression must be addressed. In many cases where grafts have failed, the causative pathology has been ischemic vasculopathy. <span style="font-size: 11pt; line-height: 115%; font-family: Arial, sans-serif; color: rgb(41, 43, 49); background-image: initial; background-position: initial; background-size: initial; background-repeat: initial; background-attachment: initial; background-origin: initial; background-clip: initial;">The biological mechanisms of the acute and chronic rejection associated with VCA, especially ischemic vasculopathy, are important areas of research. However, due to the very small number of VCA patients, the evaluation of proposed mechanisms is better addressed in an experimental model. Multiple groups have used animal models to address some of the relevant unsolved questions in VCA rejection and vasculopathy. Several model designs involving a variety of species are described in the literature. Here we present a reproducible model of VCA heterotopic hindlimb osteomyocutaneous flap in the rat that can be utilized for translational VCA research. This model allows for the serial evaluation of the graft, including biopsies and different imaging modalities, while maintaining a low level of morbidity.

Vascularized composite allograft offers life-altering benefits to transplant recipients, but the biological causes of graft rejection and vasculopathy remain poorly understood. The rodent surgical model presented here offers a reproducible, clinically relevant model of transplantation, allowing researchers to evaluate rejection events and potential therapeutic strategies to prevent their occurrence.

שונה הטרולנושא הגפיים האוסטאופהעורית מודל החולדה על השתלת ואסימטיקה מורכבת מחקר האלוגרפט

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand ...

Taking the Next Step: a Neural Coaptation Orthotopic Hind Limb Transplant Model to Maximize Functional Recovery in Rat

Limb transplant in particular and vascularized composite allotransplant (VCA) in general have wide therapeutic promise that have been stymied by current limitations in immunosuppression and functional neuromotor recovery. Many animal models have been developed for studying unique features of VCA, but here we present a robust reproducible model of orthotopic hind limb transplant in rats designed to simultaneously investigate both aspects of current VCA limitation: immunosuppression strategies and functional neuromotor recovery. At the core of the model rests a commitment to meticulous, time-tested microsurgical techniques such as hand sewn vascular anastomoses and hand sewn neural coaptation of the femoral nerve and the sciatic nerve. This approach yields durable limb reconstructions that allow for longer lived animals capable of rehabilitation, resumption of daily activities, and functional testing. With short-term treatment of conventional immunosuppressive agents, allotransplanted animals survived up to 70 days post-transplant, and isotransplanted animals provide long lived controls beyond 200 days post-operatively. Evidence of neurologic functional recovery is present by 30 days post operatively. This model not only provides a useful platform for interrogating immunological questions unique to VCA and nerve regeneration, but also allows for in vivo testing of new therapeutic strategies specifically tailored for VCA.

This protocol presents a robust, reproducible model of vascularized composite allotransplant (VCA) geared toward simultaneous study of immunology and functional recovery. The time invested in meticulous technique in a right mid-thigh hind limb orthotopic transplant with hand sewn vascular anastomoses and neural coaptation yields the ability to study functional recovery.

לוקח את הצעד הבא: מודל השתלת גפיים אחוריות קופטציה עצבית כדי למקסם את ההתאוששות תפקודית בחולדה

Limb transplant in particular and vascularized composite allotransplant (VCA) in general have wide therapeutic promise that have been stymied by current ...