Vorremmo riconoscere medica applicata per sostenere questo lavoro con e senza restrizioni borsa di studio che ci ha permesso di produrre e pubblicare il video.

<dt&gt; Porta singola nefrectomia del donatore:</dt&gt;<ol&gt;<li&gt; Il soggetto è in posizione di decubito laterale destro con il lato sinistro in alto</li&gt;<liOperativo&gt; chirurgo starà di fronte addome con il driver della fotocamera assistente in piedi a destra chirurghi e caudale</li&gt;<li&gt; Misura 5 cm su incisione ombelico sul tratto</li&gt;<li&gt; Crea incisione a ombelico ed entrare nella cavità addominale.</li&gt;<li&gt; Posizionare il Retrattore ferita Alexis</li&gt;<li&gt; Luogo porte appropriate laparoscopica al punto Seal Gel (2 porte 15 millimetri 5 millimetri e 1 porta) come disrected e mostrato in video</li&gt;<li&gt; Luogo Point sigillo Gel su Retrattore Alexis e Insuffolate l&#39;addome</li&gt;<li&gt; Mobilitare il colon discendente off del retroperitoneo</li&gt;<li&gt; Mobilitare la Milza da laterale a mediale per creare aereo tra la milza e il polo superiore del rene</li&gt;<li&gt; Dissect l&#39;uretere e la vena gonadica dal livello dei vasi iliaci fino al polo inferiore del rene</li&gt;<li&gt; Segui la Vena gonadica alla vena renale nella zona di dell&#39;ilo. La vena gonadica può essere legata e sezionata nei pressi della vena renale, se necessario</li&gt;<li&gt; Identificare e dividere le vene lombare</li&gt;<li&gt; Identificare e Dividere la vena surrenale tra i clip</li&gt;<li&gt; Dissect la vena renale circonferenza</li&gt;<li&gt; Identificare le arterie renali e sezionare la circonferenza</li&gt;<li&gt; Mobilze l&#39;intero organo off del retroperitoneo</li&gt;<li&gt; Pace a 5-12mm Port se il porto più basso 5 millimetri era stato collocato nella Seal Point Gel</li&gt;<li&gt; Legare e sezionare l&#39;uretere e la vena gonadica al Vesselswith iliaca uno Endo-GIA stapler</li&gt;<li&gt; Legare l&#39;arteria renale con un Endo-TA vascolare cucitrice e poi dividere con endoshears</li&gt;<li&gt; Legare e sezionare la vena renale con un Endo-GIA stapler</li&gt;<li&gt; Posizionare il rene in borsa e Endocatch grandi</li&gt;<li&gt; Rimozione del sigillo Point Gel e la Retrattore Alexis</li&gt;<li&gt; Rimuovi il rene dalla cavità addominale</li&gt;<li&gt; Sostituire il Seal Point Gel</li&gt;<liIndagine&gt; per l&#39;emostasi e sostituire il colon discendente nella posizione appropriata con la milza</li&gt;<li&gt; Chiudere la fascia addominale muro e poi l&#39;incisione cutanea</li&gt;<li&gt; Luogo spogliatoio</li&gt;</ol

<ol>
	<li>Canes, D., Berger, A., Aron, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparo-endoscopic+single+site+(LESS)+versus+standard+laparoscopic+left+donor+nephrectomy:+matched-pair+comparison.">Laparo-endoscopic single site (LESS) versus standard laparoscopic left donor nephrectomy: matched-pair comparison.</a> Eur Urol. 57, 95-101 (2010).</li><li>Skrekas, G., Papalois, V. E., Mitsis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+live+donor+nephrectomy:+a+step+forward+in+kidney+transplantation.">Laparoscopic live donor nephrectomy: a step forward in kidney transplantation.</a> JSLS. 7, 197-206 (2003).</li><li>Giessing, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+living-donor+nephrectomy.">Laparoscopic living-donor nephrectomy.</a> Nephrol Dial Transplant. 19, iv36-iv40 (2004).</li><li>Eng, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+laparoscopic+donor+nephrectomy+in+renal+transplantation.">The role of laparoscopic donor nephrectomy in renal transplantation.</a> Am Surg. 76, 349-353 (2010).</li><li>Kommu, S. S., Kaouk, J. H., Rane, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparo-endoscopic+single-site+surgery:+preliminary+advances+in+renal+surgery.">Laparo-endoscopic single-site surgery: preliminary advances in renal surgery.</a> BJU Int. 103, 1034-1037 (2009).</li><li>Canes, D., Desai, M. M., Aron, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transumbilical+single-port+surgery:+evolution+and+current+status.">Transumbilical single-port surgery: evolution and current status.</a> Eur Urol. 54, 1020-1029 (2008).</li><li>Desai, M. M., Berger, A. K., Brandina, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoendoscopic+single-site+surgery:+initial+hundred+patients.">Laparoendoscopic single-site surgery: initial hundred patients.</a> Urology. 74, 805-812 (2009).</li><li>Ganpule, A. P., Dhawan, D. R., Kurien, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoendoscopic+single-site+donor+nephrectomy:+a+single-center+experience.">Laparoendoscopic single-site donor nephrectomy: a single-center experience.</a> Urology. 74, 1238-1240 (2009).</li><li>Gill, I. S., Canes, D., Aron, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+port+transumbilical+(E-NOTES)+donor+nephrectomy.">Single port transumbilical (E-NOTES) donor nephrectomy.</a> J Urol. 180, 637-641 (2008).</li><li>Raman, J. D., Bensalah, K., Bagrodia, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+and+clinical+development+of+single+keyhole+umbilical+nephrectomy.">Laboratory and clinical development of single keyhole umbilical nephrectomy.</a> Urology. 70, 1039-1042 (2007).</li><li>Raybourn, J. H., Rane, A., Sundaram, 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=Laparoendoscopic+single-site+surgery+for+nephrectomy+as+a+feasible+alternative+to+traditional+laparoscopy.">Laparoendoscopic single-site surgery for nephrectomy as a feasible alternative to traditional laparoscopy.</a> Urology. 75, 100-103 (2010).</li><li>Dindo, D., Demartines, N., Clavien, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classification+of+surgical+complications:+a+new+proposal+with+evaluation+in+a+cohort+of+6336+patients+and+results+of+a+survey.+Ann+Surg.">Classification of surgical complications: a new proposal with evaluation in a cohort of 6336 patients and results of a survey. Ann Surg.</a> 240, 205-213 (2004).</li><li>Bargman, V., Sundaram, C. P., Bernie, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Randomized+trial+of+laparoscopic+donor+nephrectomy+with+and+without+hand+assistance.">Randomized trial of laparoscopic donor nephrectomy with and without hand assistance.</a> J Endourol. 20, 717-722 (2006).</li><li>El-Galley, R., Hood, N., Young, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor+nephrectomy:+A+comparison+of+techniques+and+results+of+open,+hand+assisted+and+full+laparoscopic+nephrectomy.">Donor nephrectomy: A comparison of techniques and results of open, hand assisted and full laparoscopic nephrectomy.</a> J Urol. 171, 40-43 (2004).</li><li>Velidedeoglu, E., Williams, N., Brayman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+open,+laparoscopic,+and+hand-assisted+approaches+to+live-donor+nephrectomy.">Comparison of open, laparoscopic, and hand-assisted approaches to live-donor nephrectomy.</a> Transplantation. 74, 169-172 (2002).</li><li>Raman, J. D., Bagrodia, A., Cadeddu, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-incision,+umbilical+laparoscopic+versus+conventional+laparoscopic+nephrectomy:+a+comparison+of+perioperative+outcomes+and+short-term+measures+of+convalescence.">Single-incision, umbilical laparoscopic versus conventional laparoscopic nephrectomy: a comparison of perioperative outcomes and short-term measures of convalescence.</a> Eur Urol. 55, 1198-1204 (2009).</li></ol>

Il dottor David Leeser e il dottor Giuseppe Del Pizzo sia insegnare le tecniche singola porta ai corsi sponsorizzato da Applied Medical.

Il singolo nefrectomia del donatore porta è un passo vitale successivo nell&#39;evoluzione della nefrectomia del donatore. A volte, passaggi della procedura devono essere realizzate fuori servizio a causa delle sfide inerenti alla singola porta che limita lato a lato di svincolo e della mobilità, come discusso nel video. La cosmesi è eccellente ed i pazienti ritorno alle attività molto rapidamente. Poiché la procedura si evolve nuovi strumenti saranno sviluppati che aiutante nella realizzazione di più di compiti sempre più complessi attraverso l&#39;entrata singola porta.

single port donor nephrectomy

Nel 2007, Rane ha presentato la prima nefrectomia singola porta per un piccolo mancato funzionamento dei reni al Congresso Mondiale di Endourologia. Da allora, l&#39;uso della chirurgia singola porta per la nefrectomia è espanso per includere nefrectomia del donatore. Nei prossimi due anni la tecnica fu adottata per molti altri tipi di nefrectomia per includere nefrectomia del donatore. Vi presentiamo la nostra tecnica di nefrectomia singolo donatore porta utilizzando il dispositivo Gelpoint. Abbiamo effettuato con successo questa chirurgia in oltre 100 pazienti e aggiungere questa esperienza per la nostra esperienza di oltre 1000 nefrectomia laparoscopica. Con l&#39;attrezzatura adeguata e la tecnica, nefrectomia singolo donatore porta può essere eseguita in modo sicuro ed efficace nella maggior parte dei donatori vivi. Abbiamo trovato che i nostri tempi operatori e soprattutto i nostri risultati dei trapianti non sono cambiati in modo significativo con l&#39;adozione della nefrectomia singolo donatore porta. Crediamo che solo nefrectomia donatore porto rappresenta un passo avanti nella cura dei donatori viventi.

Watch this Scientific Journal Video about Single Port Donor Nephrectomy at JoVE.com

Single Port Donor Nephrectomy

Singola porta chirurgia laparoscopica sta cambiando lo standard di cura nella cura chirurgica come niente dato che la tecnica laparoscopica è stata introdotta 20 anni fa. Vi presentiamo le tecniche di nefrectomia singolo donatore porta utilizzando il dispositivo Gelpoint. Abbiamo effettuato con successo questa chirurgia in 100 pazienti.

Porta singola donatori nefrectomia

In 2007, Rane presented the first single port nephrectomy for a small non-functioning kidney at the World Congress of Endourology.  Since that time, the ...

Research

JoVE Journal

Medicine

49.4K Views.  Weill Cornell Medical College of Cornell University. Singola porta chirurgia laparoscopica sta cambiando lo standard di cura nella cura chirurgica come niente dato che la tecnica laparoscopica è stata introdotta 20 anni fa. Vi presentiamo le tecniche di nefrectomia singolo donatore porta utilizzando il dispositivo Gelpoint. Abbiamo effettuato con successo questa chirurgia in 100 pazienti.

Video: Single Port Donor Nephrectomy

Corneal Donor Tissue Preparation for Endothelial Keratoplasty

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full thickness corneal transplantation has been the treatment to restore sight in those limited by corneal disease. Some disadvantages to this approach include a high degree of post-operative astigmatism, lack of predictable refractive outcome, and disturbance to the ocular surface. The development of Descemet's stripping endothelial keratoplasty (DSEK), transplanting only the posterior corneal stroma, Descemet's membrane, and endothelium, has dramatically changed treatment of corneal endothelial disease. DSEK is performed through a smaller incision; this technique avoids 'open sky' surgery with its risk of hemorrhage or expulsion, decreases the incidence of postoperative wound dehiscence, reduces unpredictable refractive outcomes, and may decrease the rate of transplant rejection3-6.
Initially, cornea donor posterior lamellar dissection for DSEK was performed manually1 resulting in variable graft thickness and damage to the delicate corneal endothelial tissue during tissue processing. Automated lamellar dissection (Descemet's stripping automated endothelial keratoplasty, DSAEK) was developed to address these issues. Automated dissection utilizes the same technology as LASIK corneal flap creation with a mechanical microkeratome blade that helps to create uniform and thin tissue grafts for DSAEK surgery with minimal corneal endothelial cell loss in tissue processing.
Eye banks have been providing full thickness corneas for surgical transplantation for many years. In 2006, eye banks began to develop methodologies for supplying precut corneal tissue for endothelial keratoplasty. With the input of corneal surgeons, eye banks have developed thorough protocols to safely and effectively prepare posterior lamellar tissue for DSAEK surgery. This can be performed preoperatively at the eye bank. Research shows no significant difference in terms of the quality of the tissue7 or patient outcomes8,9 using eye bank precut tissue versus surgeon-prepared tissue for DSAEK surgery. For most corneal surgeons, the availability of precut DSAEK corneal tissue saves time and money10, and reduces the stress of performing the donor corneal dissection in the operating room. In part because of the ability of the eye banks to provide high quality posterior lamellar corneal in a timely manner, DSAEK has become the standard of care for surgical management of corneal endothelial disease.
The procedure that we are describing is the preparation of the posterior lamellar cornea at the eye bank for transplantation in DSAEK surgery (Figure 1).

Endothelial corneal transplantation is a surgical technique for treatment of posterior corneal diseases. Mechanical microkeratome dissection to prepare tissue results in thinner, more symmetric grafts with less endothelial cell loss and improved outcomes. Dissections can be performed at the eye bank prior to corneal transplantation surgery.

Preparazione del tessuto corneale dei donatori per Keratoplasty Endothelial

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full ...

Ricerca

Medicina

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Utilizzando quantitativa real-time PCR per la determinazione cellulare attecchimento dei donatori in un modello murino competitivo trapianto di midollo osseo

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

5/6th Nephrectomy in Combination with High Salt Diet and Nitric Oxide Synthase Inhibition to Induce Chronic Kidney Disease in the Lewis Rat

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements of homeostasis, integrative animal models are necessary to study development and progression of CKD. To study development of CKD and novel therapeutic interventions in CKD, we use the 5/6th nephrectomy ablation model, a well known experimental model of progressive renal disease, resembling several aspects of human CKD. The gross reduction in renal mass causes progressive glomerular and tubulo-interstitial injury, loss of remnant nephrons and development of systemic and glomerular hypertension. It is also associated with progressive intrarenal capillary loss, inflammation and glomerulosclerosis. Risk factors for CKD invariably impact on endothelial function. To mimic this, we combine removal of 5/6th of renal mass with nitric oxide (NO) depletion and a high salt diet. After arrival and acclimatization, animals receive a NO synthase inhibitor (NG-nitro-L-Arginine) (L-NNA) supplemented to drinking water (20 mg/L) for a period of 4 weeks, followed by right sided uninephrectomy. One week later, a subtotal nephrectomy (SNX) is performed on the left side. After SNX, animals are allowed to recover for two days followed by LNNA in drinking water (20 mg/L) for a further period of 4 weeks. A high salt diet (6%), supplemented in ground chow (see time line Figure 1), is continued throughout the experiment. Progression of renal failure is followed over time by measuring plasma urea, systolic blood pressure and proteinuria. By six weeks after SNX, renal failure has developed. Renal function is measured using 'gold standard' inulin and para-amino hippuric acid (PAH) clearance technology. This model of CKD is characterized by a reduction in glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), hypertension (systolic blood pressure&gt;150 mmHg), proteinuria (&gt; 50 mg/24 hr) and mild uremia (&gt;10 mM). Histological features include tubulo-interstitial damage reflected by inflammation, tubular atrophy and fibrosis and focal glomerulosclerosis leading to massive reduction of healthy glomeruli within the remnant population (&lt;10%). Follow-up until 12 weeks after SNX shows further progression of CKD.

A two-stage method to establish chronic kidney disease (CKD) in the Lewis rat by surgically removing 5/6th of renal mass is described. Combination of the surgical procedure, NOS-inhibition and a high-salt diet leads to a model resembling human CKD, allowing study of causal mechanisms and development of novel therapeutic interventions.

5/6th nefrectomia in combinazione con dieta di sale e di ossido nitrico sintasi Inibizione di indurre la malattia renale cronica nel ratto Lewis

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully recapitulate early- and late-stage metastatic disease progression. Typical tumor implantation models utilize ectopic or orthotopic primary tumor implantation, but few include systemic spontaneous metastatic disease that mimics the clinical setting. This protocol describes the key steps to develop RCC disease progression stages similar to patients. First, it uses a highly metastatic mouse tumor cell line in a syngeneic model to show orthotopic tumor cell implantation. Methods include superficial and internal implantation into the sub-capsular space with cells combined with matrigel to prevent leakage and early spread. Next it describes the procedures for excision of tumor-bearing kidney (nephrectomy), with critical pre- and post- surgical mouse care. Finally, it outlines the steps necessary to monitor and assess micro-and macro-metastatic disease progression, including bioluminescent imaging as well provides a detailed visual necropsy guide to score systemic disease distribution. The goal of this protocol description is to facilitate the widespread use of clinically relevant metastatic RCC models to improve the predictive value of future therapeutic testing.&#160;

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

Models of spontaneous metastatic renal cell carcinoma (RCC) disease progression can be used for evaluating treatments in a clinically relevant setting. This protocol demonstrates different procedures for orthotopic kidney tumor cell implantation, proper nephrectomy, and finally outlines a necropsy guide for visual and bioluminescent scoring of metastatic burden and localization.

Modellazione di Carcinoma renale metastatico spontanea (MRCC) in topi dopo nefrectomia

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Corneal Donor Tissue Preparation for Descemet's Membrane Endothelial Keratoplasty

Descemet&#8217;s Membrane Endothelial Keratoplasty (DMEK) is a form of corneal transplantation in which only a single cell layer, the corneal endothelium, along with its basement membrane (Descemet&#39;s membrane) is introduced onto the recipient&#39;s posterior stroma3. Unlike Descemet&#8217;s Stripping Automated Endothelial Keratoplasty (DSAEK), where additional donor stroma is introduced, no unnatural stroma-to-stroma interface is created. As a result, the natural anatomy of the cornea is preserved as much as possible allowing for improved recovery time and visual acuity4. Endothelial Keratoplasty (EK) is the procedure of choice for treatment of endothelial dysfunction. The advantages of EK include rapid recovery of vision, preservation of ocular integrity and minimal refractive change due to use of a small, peripheral incision1. DSAEK utilizes donor tissue prepared with partial thickness stroma and endothelium. The rapid success and utilization of this procedure can be attributed to availability of eye-bank prepared precut tissue. The benefits of eye-bank preparation of donor tissue include elimination of need for specialized equipment in the operating room and availability of back up donor tissue in case of tissue perforation during preparation. In addition, high volume preparation of donor tissue by eye-bank technicians may provide improved quality of donor tissue. DSAEK may have limited best corrected visual acuity due to creation of a stromal interface between the donor and recipient cornea. Elimination of this interface with transplantation of only donor Descemet&#39;s membrane and endothelium in DMEK may improve visual outcomes and reduce complications after EK5. Similar to DSAEK, long term success and acceptance of DMEK is dependent on ease of availability of precut, eye-bank prepared donor tissue. Here we present a stepwise approach to donor tissue preparation which may reduce some barriers eye-banks face in providing DMEK grafts.

Endothelial keratoplasty is a surgical technique continuously evolving as advancements result in transplantation of thinner grafts with each succession1. Here we present a technique for controlled manual tissue dissection to allow safe and repeatable separation of endothelium and Descemet&#39;s membranes from donor corneoscleral buttons for transplantation2.

Corneale Preparazione donatore di tessuti per di Descemet Membrane Endothelial Keratoplasty

Descemet&#8217;s Membrane Endothelial Keratoplasty (DMEK) is a form of corneal transplantation in which only a single cell layer, the corneal endothelium, ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous perfusion of organs provides the opportunity to improve organ quality and allows ex situ viability assessment of donor livers prior to transplantation. This video article provides a step by step protocol for ex situ normothermic machine perfusion (37 &#176;C) of human donor livers using a device that provides a pressure and temperature controlled pulsatile perfusion of the hepatic artery and continuous perfusion of the portal vein. The perfusion fluid is oxygenated by two hollow fiber membrane oxygenators and the temperature can be regulated between 10 &#176;C and 37 &#176;C. During perfusion, the metabolic activity of the liver as well as the degree of injury can be assessed by biochemical analysis of samples taken from the perfusion fluid. Machine perfusion is a very promising tool to increase the number of livers that are suitable for transplantation.

Ex Situ Normothermic Machine Perfusion of Donor Livers

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

Ex Situ normotermica macchina di perfusione di Vivi donatori

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

Minimally Invasive Muscle Embedding (MIME) - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis

Skeletal muscle possesses regenerative capacity due to tissue-resident, muscle-fiber-generating (myogenic) satellite cells (SCs), which can form new muscle fibers under the right conditions. Although SCs can be harvested from muscle tissue and cultured in vitro, the resulting myoblast cells are not very effective in promoting myogenesis when transplanted into host muscle. Surgically exposing the host muscle and grafting segments of donor muscle tissue, or the isolated muscle fibers with their SCs onto host muscle, promotes better myogenesis compared to myoblast transplantation. We have developed a novel technique that we call Minimally Invasive Muscle Embedding (MIME). MIME involves passing a surgical needle through the host muscle, drawing a piece of donor muscle tissue through the needle track, and then leaving the donor tissue embedded in the host muscle so that it may act as a source of SCs for the host muscle. Here we describe in detail the steps involved in performing MIME in an immunodeficient mouse model that expresses a green fluorescent protein (GFP) in all of its cells. Immunodeficiency in the host mouse reduces the risk of immune rejection of the donor tissue, and GFP expression enables easy identification of the host muscle fibers (GFP+) and donor-cell-derived muscle fibers (GFP-). Our pilot data suggest that MIME can be used to implant an extensor digitorum longus (EDL) muscle from a donor mouse into the tibialis anterior (TA) muscle of a host mouse. Our data also suggest that when a myotoxin (barium chloride, BaCl2) is injected into the host muscle after MIME, there is evidence of donor-cell-derived myogenesis in the host muscle, with approximately 5%, 26%, 26% and 43% of the fibers in a single host TA muscle showing no host contribution, minimal host contribution, moderate host contribution, and maximal host contribution, respectively.

Minimally Invasive Muscle Embedding MIME - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis

We describe a novel experimental technique that we call Minimally Invasive Muscle Embedding (MIME), which is based on the evidence that skeletal muscle tissue contains viable myogenic cells that can facilitate donor-cell-mediated myogenesis when implanted into a host muscle.

Minimamente invasiva muscolo incorporamento (MIME) - una nuova tecnica sperimentale per facilitare la miogenesi donatore-cellulo-mediata

Skeletal muscle possesses regenerative capacity due to tissue-resident, muscle-fiber-generating (myogenic) satellite cells (SCs), which can form new ...

Low-Cost Single-Port (LoCoSP) Device for a Transcervical Approach in Minimally Invasive Transhiatal Esophagectomy

Esophagectomy remains the preferred option to achieve curative treatment in advanced esophageal cancer, but the choice of surgical approach remains controversial. A transthoracic approach may improve lymph node dissection, but it has considerable morbidity and respiratory complications. Transhiatal access has been demonstrated as an efficient means of minimizing post-operative complications. Minimally invasive transhiatal esophagectomy may reduce operative trauma and morbidity as well as enhance postoperative recovery with no compromise in cancer recurrence or survival. Nevertheless, it has a technical limitation in terms of cervical esophagus dissection. Thus, a low-cost single-port device was developed to complete upper mediastinal dissection by a transcervical approach during minimally invasive transhiatal esophagectomy. This device uses a nasogastric tube, a number-eight sterile glove, a sterile sponge, and 3 permanent 5-mm trocars. The step-by-step process of transhiatal esophagectomy and the development of this device are described. This technique allows for the dissection of the upper mediastinum, as well as the esophagus over the aortic arch and behind the superior portion of the trachea. The harvesting of lymph nodes along the left recurrent laryngeal nerve and paratracheals was improved.

Low-Cost Single-Port LoCoSP Device for a Transcervical Approach in Minimally Invasive Transhiatal Esophagectomy

Here, we describe a step-by-step description of transhiatal esophagectomy and the development of a low-cost single-port device for a transcervical approach in a minimally invasive transhiatal esophagectomy.

Dispositivo a porta singola (LoCoSP) a basso costo per un approccio transcervicale nell'esofagectomia transiatale minimamente invasiva

Esophagectomy remains the preferred option to achieve curative treatment in advanced esophageal cancer, but the choice of surgical approach remains ...

Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years confirm that the outcomes after lung transplantation from cDCD are similar to those from brain death donors; however, the utilization of lungs from asystole donors remains low. Several reasons may be involved: different legal frameworks among countries and centers with different premortem interventions, inadequate lung donor care before procurement, or even poor experience with cDCD procedures and protocols.
Initially, the rapid recovery technique was commonly employed for the procurement of thoracic and abdominal organs in cDCD, but, in the last decade, abdominal normothermic regional perfusion (ANRP) with extracorporeal membrane oxygenation devices has become a useful method to restore blood flow to abdominal organs, allowing their quality improvement and their functional assessment prior to transplantation. This makes the donation procedure more complex and generates doubts about injury to the grafts due to dual temperature.
The aim of this article is to describe a protocol based on a single center experience with Maastricht III donors combining lung cooling rapid recovery in the thorax and abdominal normothermic regional perfusion. Tips and tricks focused on premortem interventions and lung procurement procedure techniques are explained. This may help to minimize the reluctance among professionals to use this combined technique and encourage other donor centers to use it, despite the increased complexity of the procedure.

The protocol combines a lung cooling rapid recovery technique with abdominal normothermic regional perfusion for abdominal graft procurement in controlled asystole donors, which is a safe and useful method to expand the donor pool.

Approvvigionamento di recupero rapido polmonare combinato con perfusione regionale normomica addominale nella donazione controllata dopo morte circolatoria

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years ...

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

Trapianto di fegato suino senza bypass veno-venoso come modello di donatore a criteri estesi

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...