Bu çalışma NIH hibe R01AI077610 tarafından finanse edildi.

Tüm işlemler Bakım ve Ulusal Sağlık Enstitüsü (NIH) Laboratuvar Hayvanları Kullanım Kılavuzu uyarınca yapılmıştır ve Johns Hopkins Üniversitesi Hayvan Bakım ve Kullanım Komitesi (JHUACUC) tarafından onaylanmıştır. Belirli uygulama usulleri ACUC protokolleri MO13M292 ve MO13M370 altında yapıldı. 1. Donör Cilt Hasat <ol><li> (- Fare konisi ile 2% 1 bakım,% 4 indüksiyon buharlaştırıcı) izofluran ile donör fare anestezisi. anestezi derinliği izlemek için ayak tutam çekilme refleksi kullanın. </li><li> Bir elektrikli tıraş makinesi kullanarak, hayvanın arka tıraş ve (örneğin, Betadine)% 10 povidon iyot ile dezenfekte edin. </li><li> areolar bağ dokusu düzeyinde künt diseksiyon ile steril makas, eldiven ve aseptik teknik, boyun kalça hasat donör arka cilt, kullanma. </li><li> hasat sonrası servikal dislokasyon ile hayvan Euthanizederi grefti. </li><li> Mikroskop altında, ince tenotomi makas kullanarak geri deriden bağ dokusu, yağ dokusu ve panniculus carnosus ayırın. panniculus carnosus cilt seğirmesi hareketleri sorumlu ince, şeffaf bir kastır. </li><li> Steril aletler tekniği kullanarak, 15 mm x 15 mm greft yatağına 10 mm x 10 mm geri deriden 15 mm x 15 mm greft kesip. </li><li> Buz üzerinde bir petri steril fosfat tamponlu salin (PBS) ile ıslatılmış etme yöntemleriyle ilgili greft saklayın. Not: 8-10 greft, bir verici bir fare elde edilebilir. </li></ol> 2. Alıcı Cilt Nakli <ol><li> (- Fare konisi ile 2% 1 bakım,% 4 indüksiyon buharlaştırıcı) izofluran ile alıcı fare anestezisi. anestezi derinliği izlemek için ayak tutam çekilme refleksi kullanın. </li><li> Ameliyat sonrası ağrı tedavisi için Buprenorfin 0.02 mg / kg BW yönetme. </li><li> Hayvanın arka tarafı Tıraş nerede greft will takılı olması ve% 10 povidon iyot ile dezenfekte edin. </li><li> makas kullanarak, derinin x 15 mm kare 15-mm 10 mm x 10 mm&#39;lik kesti. kusur boyutu greft daha (% 10) biraz daha büyük olması gerekir. kadar yüzeysel mümkün olduğunca kesin. panniculus carnosus ve damarları korumak için özen gösterin. panniculus carnosus onun hareketlilik ve buna yüzeysel koşmak kan damarları tarafından altta yatan fasya ayırt olabilir. </li><li> kenarları boyunca kıvrımlar kaçınarak, greft yatakta greft yerleştirin. </li><li> Köşelerde ve her kenarın ortasında 8 sütür yerleştirin. Her sütür için, çevredeki alıcı deri altına yatak greft panniculus carnosus yoluyla daha sonra greft içinden iğne geçmesi ve. </li><li> anestezi maskesi çıkarın ve hayvan yapışkan bandaj uygulamadan önce anestezi kısmen kurtarmak edelim. </li><li> greft üzerine katlanmış gazlı bez ile bir yapışkan bandaj alıcı fare sarın. kesme, iki bandaj birleştirerek bandaj yapmakyapışkan birinin parçası ve birlikte iki emici pedler yerleştirerek. </li><li> bandaj toraks gezi kısıtlayan ve nefes olmadığından emin olmak için kurtarma sırasında yakından fare izleyin. solunum sayısı azalır ya da hayvan sığ gasp ya da nefes başlarsa bandajı çıkarın. </li></ol> 3. Ameliyat Sonrası Bakım <ol><li> Enfeksiyon profilaksisi için ameliyattan sonra enrofloxacin 5 mg / kg yönetmek. </li><li> tamamen anestezi kurtarır kadar mikrodalga ısıtma yastığı üzerinde temiz bir kafes içinde nakledilen fare yerleştirin. </li><li> konut tesisine iade postoperatif önce 1 saat için fare gözlemleyin. </li><li> Yedi gün ameliyat sonrası Adım 2.1 gibi fare uyutmak. bandaj ventral tarafta boyunca sadece keserek bandaj çıkarın. </li><li> Gözlemlemek ve kabuklanma, kasılma veya sertlik belirtileri ertesi gün greft palpe. Varsa, greft uygun vasculariza elde olmayabiliryon ve teknik bir arıza düşünülmelidir. </li><li> ret belirtileri günlük izleyin. greft dokusunun ≥90% nekrotik hale geldiğinde reddedilen greft düşünün. </li><li> reddedilen hayvanları euthanize ve analiz için onları hasat. </li></ol>

<ol>
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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+Hind+Limb+Transplantation+in+the+Mouse.">Orthotopic Hind Limb Transplantation in the Mouse.</a> J Vis Exp. , (2016).</li><li>Oh, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Microsurgical+Model+for+Heterotopic,+En+Bloc+Chest+Wall,+Thymus,+and+Heart+Transplantation+in+Mice.">A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice.</a> J Vis Exp. , (2016).</li><li>Oberhuber, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+cervical+heart+transplantation+model+using+a+modified+cuff+technique.">Murine cervical heart transplantation model using a modified cuff technique.</a> J Vis Exp. , e50753 (2014).</li><li>Jones, T. R., Shirasugi, N., Adams, A. B., Pearson, T. C., Larsen, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravital+microscopy+identifies+selectins+that+regulate+T+cell+traffic+into+allografts.">Intravital microscopy identifies selectins that regulate T cell traffic into allografts.</a> J Clin Invest. 112, 1714-1723 (2003).</li><li>Celli, S., Albert, M. L., Bousso, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+the+innate+and+adaptive+immune+responses+underlying+allograft+rejection+by+two-photon+microscopy.">Visualizing the innate and adaptive immune responses underlying allograft rejection by two-photon microscopy.</a> Nat Med. 17, 744-749 (2011).</li><li>Medawar, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+behaviour+and+fate+of+skin+autografts+and+skin+homografts+in+rabbits:+A+report+to+the+War+Wounds+Committee+of+the+Medical+Research+Council.">The behaviour and fate of skin autografts and skin homografts in rabbits: A report to the War Wounds Committee of the Medical Research Council.</a> J Anat. 78, 176-199 (1944).</li><li>Billingham, R. E., Brent, L., Medawar, P. B., Sparrow, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+studies+on+tissue+transplantation+immunity.+I.+The+survival+times+of+skin+homografts+exchanged+between+members+of+different+inbred+strains+of+mice.">Quantitative studies on tissue transplantation immunity. I. The survival times of skin homografts exchanged between members of different inbred strains of mice.</a> Proc R Soc Lond B Biol Sci. 143, 43-58 (1954).</li><li>Garrod, K. R., Cahalan, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+skin+transplantation.">Murine skin transplantation.</a> J Vis Exp. , (2008).</li><li>Schmaler, M., Broggi, M. A., Rossi, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+tail+skin+to+study+allogeneic+CD4+T+cell+responses+in+mice.">Transplantation of tail skin to study allogeneic CD4 T cell responses in mice.</a> J Vis Exp. , e51724 (2014).</li><li>Bergstresser, P. R., Toews, G. B., Gilliam, J. N., Streilein, J. 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Yazarlar ifşa hiçbir şey yok.

Tavşan ve farelerde daha sonra insan çalışmalarında ilk ve Medawar tarafından piyasaya sunulmasından bu yana, deri transplantasyonu allojenik bağışıklık tepkilerinin 6,7 çalışmaları için paha biçilmez bir model olmuştur. Bu yazıda, üst ve alt sırt derisi kullanılarak büyük ölçekli, non-vaskülarize, tam kalınlıkta deri nakli bir model sunuyoruz. Kuyruk deri veya Nakledilecek doku kaynağı olarak fare kulak derisi kullanılarak da dahil olmak üzere çeşitli alternatif yöntemler...

Organ nakli son dönem organ yetmezliği olan hastalarda tercih edilen tedavi olup, sonuçlar cerrahi işlemler ve immünsüpresyondaki gelişmeler ile son derece düzeldi. Ancak, uzun süreli immünsüpresyon önemli yan etkileri ile ilişkilidir ve hoşgörüyü teşvik yeni stratejilerin geliştirilmesi, modern nakli araştırmanın amacı kalır. Çeşitli hayvan modelleri allograft reddinin mekanizmaları incelemek ve immünosupresyon aşı reddinin engellenmesi için ve uzun vadeli dayanıklılık 1-3 teşvik etmek için yaklaşımlar test etmek için, transplantasyon temel araştırma için geliştirilmiştir. Fare modelleri nedeniyle tanı ve tedavi antikorlar ve iyi tanımlanmış kendilenmiş ve transgenik suşlar seçkin ve geniş müsaitlik durumuna immünolojik araştırmaların temelini haline gelmiştir. Cilt nakli özel mikrocerrahi beceri gerektiren ve can değil basit bir işlemdirKolayca postoperatif izlenecektir. Birlikte ele alındığında, fare deri nakli greft reddi 4,5 sırasında antijen sunumu, hücre kaçakçılığı ve doku yıkımı da dahil olmak üzere immün yanıt katılan birçok açıdan incelemek için olağanüstü bir araç olmuştur. Burada, fare modeli kullanılarak tam kalınlıkta deri transplantasyonu için adım-adım prosedürü göstermek ve biz nakledilen deri başarılı bütünleşme için gerekli önemli adımlar açıklanmaktadır.

<table><tbody><tr><td>Straight micro forceps</td><td>Sigma</td><td>F4017</td><td></td></tr><tr><td>Curved micro forceps</td><td>Aesculap</td><td>BD333R</td><td></td></tr><tr><td>Curved Stevens tenotomy scissors</td><td>Aesculap</td><td>BC905R</td><td></td></tr><tr><td>Mayo dissecting scissors</td><td>Sigma</td><td>S3146</td><td></td></tr><tr><td>Micro needle holder</td><td>Aesculap</td><td>BM563R</td><td></td></tr><tr><td>Sterile gauze</td><td>Covidien</td><td>441218</td><td></td></tr><tr><td>6-0 Nylon suture</td><td>MWI</td><td>31849</td><td></td></tr><tr><td>Plastic Strips Band&amp;#45;Aid</td><td>Johnson &amp;amp; Johnson</td><td></td><td>Obtained from pharmacy</td></tr><tr><td>10 cm Petri dish</td><td>Fisherbrand</td><td>FB0875712</td><td></td></tr><tr><td>PBS</td><td>Quality Biological</td><td>119069131</td><td></td></tr><tr><td>Buprenorphine</td><td></td><td></td><td>DEA Number required; Obtained from hospital pharmacy</td></tr><tr><td>Enrofloxacin</td><td>Bayer Health Care</td><td>186599</td><td></td></tr></tbody></table>

murine full-thickness skin transplantation

Mürin tam kalınlıkta deri transplantasyonu alloimmun tepki ve greft reddini incelemek için iyi kurulmuş bir in vivo modeldir. insanlara sınırlı uygulanmasına rağmen, farelerde deri transplantasyon yaygın Nakil için kullanılmıştır. Prosedür öğrenmek ve kolay uygulanabilir olması ve hassas mikrocerrahi teknikler de kapsamlı bir eğitim gerektirmez. Ayrıca, bu modelde graft reddi çok tekrarlanabilir immünolojik reaksiyon meydana gelir ve kolayca doğrudan kontrol ve elle muayene ile izlenir. Buna ek olarak, donör uyumlu veya üçüncü taraf deri grefti ile ikincil deri nakli alternatif ve donör spesifik tolerans değerlendirmek için basit bir yöntem olarak daha karmaşık nakli modellerinde yapılabilir. komplikasyonlar düşük ve işlem sonrası anestezi doz aşımı ya da solunum sıkıntısı ile sınırlı genel olarak. Greft yetmezliği, diğer taraftan, gr yetersiz hazırlık sonucu sık görülürgreft yatakta kıç, yanlış konumlandırma, ya da bandaj uygunsuz yerleştirme. Bu yazıda, farelerde tam kalınlıkta deri transplantasyonu için bir protokol mevcut ve başarılı bir işlem için gerekli olan önemli adımları açıklar.

Alıcı fare üzerindeki bandajın yerleştirme işleminin önemli bir adımdır. Deri grefti omuz, kalça, omurga ve (Şekil 1) arasında, alıcı gövde üzerinde konumlandırılmış. Bandaj katlanmış gazlı bez ve iki plastik yapışkanlı bandaj kombinasyonu ile yapılır. Alıcı fare bandajın ortasına gazlı bez üzerinde aşağı greft ile yerleştirilir. iki kavisli mikro forseps kullanılarak, bandajın alt ucu, ilk çekilir ve daha sonra bir bandaj üst ka...

Watch this Scientific Journal Video about Murine Full-thickness Skin Transplantation at JoVE.com

Murine Full-thickness Skin Transplantation

Mürin tam kalınlıkta deri transplantasyonu alloimmun ortamda reddini incelemek için iyi kurulmuş bir modeldir. &gt; C57BL / 6 tam kalınlıkta deri nakli - Burada bir BALB / c performans katılan her adımda bir öğretici sağlar.

Mürin Tam kat Cilt Nakli

Murine full-thickness skin transplantation is a well-established in vivo model to study alloimmune response and graft rejection. Despite its limited ...

murine-full-thickness-skin-transplantation

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

20.0K Görüntüleme.  Johns Hopkins University School of Medicine.  Mürin tam kalınlıkta deri transplantasyonu alloimmun ortamda reddini incelemek için iyi kurulmuş bir modeldir. > C57BL / 6 tam kalınlıkta deri nakli - Burada bir BALB / c performans katılan her adımda bir öğretici sağlar. 

Video: Murine Full-thickness Skin Transplantation

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells behave differently when they are grown within a 3D extra-cellular matrix and also interact with other cells (1-5). Mouse models have been broadly utilized to study tissue morphogenesis in vivo. However mouse and human skin have significant differences in cellular architecture and physiology, which makes it difficult to extrapolate mouse studies to humans. Since melanocytes in mouse skin are mostly localized in hair follicles, they have distinct biological properties from those of humans, which locate primarily at the basal layer of the epidermis. The recent development of 3D human skin reconstruct models has enabled the field to investigate cell-matrix and cell-cell interactions between different cell types. The reconstructs consist of a "dermis" with fibroblasts embedded in a collagen I matrix, an "epidermis", which is comprised of stratified, differentiated keratinocytes and a functional basement membrane, which separates epidermis from dermis. Collagen provides scaffolding, nutrient delivery, and potential for cell-to-cell interaction. The 3D skin models incorporating melanocytic cells recapitulate natural features of melanocyte homeostasis and melanoma progression in human skin. As in vivo, melanocytes in reconstructed skin are localized at the basement membrane interspersed with basal layer keratinocytes. Melanoma cells exhibit the same characteristics reflecting the original tumor stage (RGP, VGP and metastatic melanoma cells) in vivo. Recently, dermal stem cells have been identified in the human dermis (6). These multi-potent stem cells can migrate to the epidermis and differentiate to melanocytes.

In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.

Üç Boyutlu İnsan Cilt Model Yeniden İnşa: Aracı Normal Cilt ve Melanom İlerleme Eğitim

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

Biyomühendislik

Murine Skin Transplantation

As one of the most stringent and least technically challenging models, skin transplantation is a standard method to assay host T cell responses to MHC-disparate donor antigens. The aim of this video-article is to provide the viewer with a step-by-step visual demonstration of skin transplantation using the mouse model. The protocol is divided into 5 main components: 1) harvesting donor skin; 2) preparing recipient for transplant; 3) skin transplant; 4) bandage removal and monitoring graft rejection; 5) helpful hints. Once proficient, the procedure itself should take &lt;10 min to perform.

Allogeneic skin transplantation is a standard model to assay host T cell responses to MHC-disparate donor antigens. This video-article provides a visual tutorial of each step involved in performing a BALB/c--&gt;C57BL/6 skin transplant.

Murin Deri nakli

As one of the most stringent and least technically challenging models, skin transplantation is a standard method to assay host T cell responses to ...

Biyoloji

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to evaluate new strategies that aim at improving vascularization, reliable methods are required to make the in-growth of new blood vessels into bio-artificial scaffolds visible and quantify the results. Over the past couple of years, our group has introduced a full skin defect model that enables the direct visualization of blood vessels by transillumination and provides the possibility of quantification through digital segmentation. In this model, one surgically creates full skin defects in the back of mice and replaces them with the material tested. Molecules or cells of interest can also be incorporated in such materials to study their potential effect. After an observation time of one&#8217;s own choice, materials are explanted for evaluation. Bilateral wounds provide the possibility of making internal comparisons that minimize artifacts among individuals as well as of decreasing the number of animals needed for the study. In comparison to other approaches, our method offers a simple, reliable and cost effective analysis. We have implemented this model as a routine tool to perform high-resolution screening when testing vascularization of different biomaterials and bio-activation approaches.

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Vascularization is key to approaches in successful tissue engineering. Therefore, reliable technologies are required to evaluate the development of vascular networks in tissue-constructs. Here we present a simple and cost-effective method to visualize and quantify vascularization in vivo.

A Tam Cilt Hata Modeli Biyomateryallerin vaskülarizasyonunu değerlendirin<em&gt; In Vivo</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Yara Şifa murin Modeli

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Tıp

Minced Tissue in Compressed Collagen: A Cell-containing Biotransplant for Single-staged Reconstructive Repair

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped human cell culture facilities. These methods include isolation of cells in single cell suspension and several laborious and time-consuming events before transplantation back to the patient. Previous studies suggest that the body itself could be used as a bioreactor for cell expansion and regeneration of tissue in order to minimize ex vivo manipulations of tissues and cells before transplanting to the patient. The aim of this study was to demonstrate a method for tissue harvesting, isolation of continuous epithelium, mincing of the epithelium into small pieces and incorporating them into a three-layered biomaterial. The three-layered biomaterial then served as a delivery vehicle, to allow surgical handling, exchange of nutrition across the transplant, and a controlled degradation. The biomaterial consisted of two outer layers of collagen and a core of a mechanically stable and slowly degradable polymer. The minced epithelium was incorporated into one of the collagen layers before transplantation. By mincing the epithelial tissue into small pieces, the pieces could be spread and thereby the propagation of cells was stimulated. After the initial take of the transplants, cell expansion and reorganization would take place and extracellular matrix mature to allow ingrowth of capillaries and nerves and further maturation of the extracellular matrix. The technique minimizes ex vivo manipulations and allow cell harvesting, preparation of autograft, and transplantation to the patient as a simple one-stage intervention. In the future, tissue expansion could be initiated around a 3D mold inside the body itself, according to the specific needs of the patient. Additionally, the technique could be performed in an ordinary surgical setting without the need for sophisticated cell culturing facilities.

Tissue engineering often includes in vitro expansion in order to create autografts for tissue regeneration. In this study a method for tissue expansion, regeneration, and reconstruction in vivo was developed in order to minimize the processing of cells and biological materials outside the body.

Sıkıştırılmış Kolajen kıyılmış Doku: Tek aşamalı Rekonstrüktif Onarım için Cep içeren Biotransplant

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped ...

Transplantation of Tail Skin to Study Allogeneic CD4 T Cell Responses in Mice

The study of T cell responses and their consequences during allo-antigen recognition requires a model that enables one to distinguish between donor and host T cells, to easily monitor the graft, and to adapt the system in order to answer different immunological questions. Medawar and colleagues established allogeneic tail-skin transplantation in mice in 1955. Since then, the skin transplantation model has been continuously modified and adapted to answer specific questions. The use of tail-skin renders this model easy to score for graft rejection, requires neither extensive preparation nor deep anesthesia, is applicable to animals of all genetic background, discourages ischemic necrosis, and permits chemical and biological intervention. 
In general, both CD4+ and CD8+ allogeneic T cells are responsible for the rejection of allografts since they recognize mismatched major histocompatibility antigens from different mouse strains. Several models have been described for activating allogeneic T cells in skin-transplanted mice. The identification of major histocompatibility complex (MHC) class I and II molecules in different mouse strains including C57BL/6 mice was an important step toward understanding and studying T cell-mediated alloresponses. In the tail-skin transplantation model described here, a three-point mutation (I-Abm12) in the antigen-presenting groove of the MHC-class II (I-Ab) molecule is sufficient to induce strong allogeneic CD4+ T cell activation in C57BL/6 mice. Skin grafts from I-Abm12 mice on C57BL/6 mice are rejected within 12-15 days, while syngeneic grafts are accepted for up to 100 days. The absence of T cells (CD3-/- and Rag2-/- mice) allows skin graft acceptance up to 100 days, which can be overcome by transferring 2 x 104 wild type or transgenic T cells. Adoptively transferred T cells proliferate and produce IFN-&#947; in I-Abm12-transplanted Rag2-/- mice.

Tail-skin transplantation is a powerful model for studying T cell-dependent rejection and tolerance induction during allogeneic immune responses in mice. The advantages of this protocol are minor invasive surgery, and ease of monitoring with no need to sacrifice the recipient mouse.

Farelerde allojenik CD4 + T hücre Cevapları Eğitim için Kuyruk Cilt Nakli

The study of T cell responses and their consequences during allo-antigen recognition requires a model that enables one to distinguish between donor and ...

İmmünoloji ve Enfeksiyon

Generation of 3D Skin Organoid from Cord Blood-derived Induced Pluripotent Stem Cells

The skin is the body&#8217;s largest organ and has many functions. The skin acts as a physical barrier and protector of the body and regulates bodily functions. Biomimetics is the imitation of the models, systems, and elements of nature for the purpose of solving complex human problems1. Skin biomimetics is a useful tool for in vitro disease research and in vivo regenerative medicine. Human induced pluripotent stem cells (iPSCs) have the characteristic of unlimited proliferation and the ability of differentiation to three germ layers. Human iPSCs are generated from various primary cells, such as blood cells, keratinocytes, fibroblasts, and more. Among them, cord blood mononuclear cells (CBMCs) have emerged as an alternative cell source from the perspective of allogeneic regenerative medicine. CBMCs are useful in regenerative medicine because human leukocyte antigen (HLA) typing is essential to the cell banking system. We provide a method for the differentiation of CBMC-iPSCs into keratinocytes and fibroblasts and for generation of a 3D skin organoid. CBMC-iPSC-derived keratinocytes and fibroblasts have characteristics similar to a primary cell line. The 3D skin organoids are generated by overlaying an epidermal layer onto a dermal layer. By transplanting this 3D skin organoid, a humanized mice model is generated. This study shows that a 3D human iPSC-derived skin organoid may be a novel, alternative tool for dermatologic research in vitro and in vivo.

We propose a protocol that shows how to differentiate induced pluripotent stem cell-derived keratinocytes and fibroblasts and generate a 3D skin organoid, using these keratinocytes and fibroblasts. This protocol contains an additional step of generating a humanized mice model. The technique presented here will improve dermatologic research.

3B cilt Organoid kablosu kan kaynaklı nesil Pluripotent kök hücreler indüklenen

The skin is the body&#8217;s largest organ and has many functions. The skin acts as a physical barrier and protector of the body and regulates bodily ...

Gelişim Biyolojisi

Xenograft Skin Model to Manipulate Human Immune Responses In Vivo

The human skin xenograft model, in which human donor skin is transplanted onto an immunodeficient mouse host, is an important option for translational research in skin immunology. Murine and human skin differ substantially in anatomy and immune cell composition. Therefore, traditional mouse models have limitations for dermatological research and drug discovery. However, successful xenotransplants are technically challenging and require optimal specimen and mouse graft site preparation for graft and host survival. The present protocol provides an optimized technique for transplanting human skin onto mice and discusses necessary considerations for downstream experimental aims. This report describes the appropriate preparation of a human donor skin sample, assembly of a surgical setup, mouse and surgical site preparation, skin transplantation, and post-surgical monitoring. Adherence to these methods allows for maintenance of xenografts for over 6 weeks post-surgery. The techniques outlined below allow maximum grafting efficiency due to the development of engineering controls, sterile technique, and pre- and post-surgical conditioning. Appropriate performance of the xenograft model results in long-lived human skin graft samples for experimental characterization of human skin and preclinical testing of compounds in vivo.

Xenograft Skin Model to Manipulate Human Immune Responses In Vivo

The present protocol describes how to graft human skin onto non-obese diabetic (NOD)-scid interleukin-2 gamma chain receptor (NSG) mice. A detailed description of the preparation of human skin for transplant, preparation of mice for transplant, transplantation of split-thickness human skin, and post-transplantation recovery procedure are included in the report.

İnsan Bağışıklık Tepkilerini In Vivo'da Manipüle Etmek için Ksenograft Cilt Modeli

The human skin xenograft model, in which human donor skin is transplanted onto an immunodeficient mouse host, is an important option for translational ...

A Murine Model of a Burn Wound Reconstructed with an Allogeneic Skin Graft

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and require surgical debridement and skin grafting. Successful integration of the donor graft into a recipient wound bed depends on timely recruitment of immune cells, robust angiogenic response and new extracellular matrix formation. The development of novel therapeutic agents, which target some key processes involved in wound healing, are hindered by the lack of reliable preclinical models with optimized objective assessment of wound closure. Here, we describe an inexpensive and reproducible model of experimental full thickness burn wound reconstructed with an allogeneic skin graft. The wound is induced on the dorsum surface of anaesthetized inbred wild type mice from the BALB/C and SKH1-Hrhr backgrounds. The burn is produced using a brass template measuring 10 mm in diameter, which is preheated to 80 &#176;C and delivered at a constant pressure for 20 s. Burn eschar is excised 24 hours after the injury and replaced with a full thickness graft harvested from the tail of a genetically similar donor mouse. No specialized equipment is required for the procedure and surgical techniques are straightforward to follow. The method may be effortlessly implemented and reproduced in most research settings. Certain limitations are associated with the model. Due to technical difficulties, the harvest of thinner split thickness skin grafts is not possible. The surgical method we describe here allows for the reconstruction of burn wounds using full thickness skin grafts. It may be used to carry out preclinical therapeutic testing.

The aim of this study was to develop a murine model of burn wound healing. A thermal burn was induced on the dorsal skin of mice using a preheated brass template. Burned tissue was debrided and overlaid with a skin graft harvested from the tail of a genetically similar donor mouse.

Allojenik Deri Grefti ile Yeniden Yapılandırılan Yanık Yarasının Murine Modeli

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and ...

A Technique for Human Skin Grafting in a Mouse Model

Source: Moss, M. I. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/64040">Xenograft Skin Model to Manipulate Human Immune Responses&#160;In Vivo.</a>&#160;J. Vis. Exp. (2022)
This video demonstrates a method for grafting human skin onto an immunodeficient mouse with a suppressed neutrophil response. The method involves applying human split-thickness skin to the mouse&#39;s graft bed, sealing it, and covering it for recovery. Eventually, the human skin adheres to the mouse&#39;s skin, establishing vascularization and circulation, resulting in a successful skin xenograft.

Bir fare modelinde insan derisi grefti için bir teknik

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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