Advances in single-cell sequencing: insights from organ transplantation.

Single-cell RNA sequencing (scRNA-seq) is a comprehensive technical tool to analyze intracellular and intercellular interaction data by whole transcriptional profile analysis. Here, we describe the application in biomedical research, focusing on the immune system during organ transplantation and rejection. Unlike conventional transcriptome analysis, this method provides a full map of multiple cell populations in one specific tissue and presents a dynamic and transient unbiased method to explore the progression of allograft dysfunction, starting from the stress response to final graft failure. This promising sequencing technology remarkably improves individualized organ rejection treatment by identifying decisive cellular subgroups and cell-specific interactions.

Improving the Rate of Translation of Tissue Engineering Products.

Over 100 000 research articles and 9000 patents have been published on tissue engineering (TE) in the past 20 years. Yet, very few TE products have made their way to the market during the same period. Experts have proposed a variety of strategies to address the lack of translation of TE products. However, since these proposals are guided by qualitative insights, they are limited in scope and impact. Machine learning is utilized in the current study to analyze the entire body of patents that have been published over the past twenty years and understand patenting trends, topics, areas of application, and exemplifications. This analysis yields surprising and little-known insights about the differences in research priorities and perceptions of innovativeness of tissue engineers in academia and industry, as well as aids to chart true advances in the field during the past twenty years. It is hoped that this analysis and subsequent proposal to improve translational rates of TE products will spur much needed dialogue about this important pursuit.

Strategies based on organ decellularization and recellularization.

Transplantation is the only curative treatment option available for patients suffering from end-stage organ failure, improving their quality of life and long-term survival. However, because of organ scarcity, only a small number of these patients actually benefit from transplantation. Alternative treatment options are needed to address this problem. The technique of whole-organ decellularization and recellularization has attracted increasing attention in the last decade. Decellularization includes the removal of all cellular components from an organ, while simultaneously preserving the micro and macro anatomy of the extracellular matrix. These bioscaffolds are subsequently repopulated with patient-derived cells, thus constructing a personalized neo-organ and ideally eliminating the need for immunosuppression. However, crucial problems have not yet been satisfyingly addressed and remain to be resolved, such as organ and cell sources. In this review, we focus on the actual state of organ de- and recellularization, as well as the problems and future challenges.

La inserción del enfermero en la comisión intrahospitalaria de donación de órganos y tejidos / A inserção do enfermeiro em comissão intra-hospitalar de doação de órgãos e tecidos / The insertion of the nurse in intra-hospital commission of organ and tissue donation

Objetivo: Conocer cómo es la inserción del enfermero en la comisión intrahospitalaria de donación de órganos y tejidos para trasplante, así como la adquisición de conocimientos para actuar en ella. Método: Investigación cualitativa, descriptiva y exploratoria, realizada con 12 enfermeros de la comisión intrahospitalaria de donación de órganos y tejidos. Los datos fueron recolectados a través de entrevistas semiestructuradas y analizados mediante análisis de contenido. Resultados: Mostraron que los enfermeros son señalados para trabajar en la comisión intrahospitalaria de donación de órganos y tejidos para trasplante sin la preparación adecuada. El conocimiento se produce después de la inclusión en la referida comisión, normalmente con los eventos y apoyo en la literatura de área, ya que no existe un enfoque consistente sobre el tema en la formación profesional. Conclusión: Las medidas educativas deben ser incluidas en la academia y en los servicios de salud, con el fin de proporcionar los apoyos necesarios para que el enfermero puedan tener una participación más efectiva en estas comisiones

Objetivo: conhecer como se dá a inserção do enfermeiro em comissão intra-hospitalar de doação de órgãos e tecidos para transplante, bem como a obtenção de conhecimento para atuação nesta. Método: pesquisa qualitativa, descritiva e exploratória, realizada com 12 enfermeiros de comissão intra-hospitalar de doação de órgãos e tecidos. Os dados foram coletados por meio de entrevista semiestruturada e analisados através de análise de conteúdo. Resultados: evidenciou-se que os enfermeiros são indicados para atuar em comissão intra-hospitalar de doação de órgãos e tecidos para transplante, sem o preparo adequado. O conhecimento ocorre posteriormente à inserção na referida comissão, normalmente, junto a eventos e apoio na literatura da área, já que não há abordagem consistente sobre o tema na formação profissional. Conclusão: medidas educativas devem ser inseridas na academia e serviços de saúde, como forma de prover subsídios necessários para que o enfermeiro possa ter uma participação mais efetiva nessas comissões

Objective: To recognize how the insertion of the nurse in an in-hospital commission of donation of organs and tissues for transplantation is, as well as the obtaining of knowledge to act in this one. Method: It is a qualitative, descriptive and exploratory research, carried out with 12 nurses of intra-hospital commission of organ and tissue donation. Data were collected through a semi-structured interview and analyzed through content analysis. Results: It was evidenced that the nurses are indicated to act in intra-hospital commission of donation of organs and tissues for transplantation without adequate preparation. The knowledge occurs later to the insertion in the referred committee, usually, next to events and support in the literature of the area, since there is no consistent approach about the subject in the professional formation. Conclusion: Educational measures should be included in the health services and academia as a way of providing the necessary subsidies so that nurses can have a more effective participation in these commissions

Bio-scaffolds in organ-regeneration: Clinical potential and current challenges.

Cadaveric organ transplantation represents the definitive treatment option for end-stage disease but is restricted by the shortage of clinically-viable donor organs. This limitation has, in part, driven current research efforts for in vitro generation of transplantable tissue surrogates. Recent advances in organ reconstruction have been facilitated by the re-purposing of decellularized whole organs to serve as three-dimensional bio-scaffolds. Notably, studies in rodents indicate that such scaffolds retain native extracellular matrix components that provide appropriate biochemical, mechanical and physical stimuli for successful tissue/organ reconstruction. As such, they support the migration, adhesion and differentiation of reseeded primary and/or pluripotent cell populations, which mature and achieve functionality through short-term conditioning within specialized tissue bioreactors. Whilst these findings are encouraging, significant challenges remain to up-scale the present technology to accommodate human-sized organs and thereby further the translation of this approach towards clinical use. Of note, the diverse structural and cellular composition of large mammalian organ systems mean that a "one-size fits all" approach cannot be adopted either to the methods used for their decellularization or the cells required for subsequent re-population, to create fully functional entities. The present review seeks to highlight the clinical potential of decellularized organ bio-scaffolds as a route to further advance the field of tissue- and organ-regeneration, and to discuss the challenges which are yet to be addressed if such a technology is ever to become a credible rival to conventional organ allo-transplantation.

Tracheal Replacement Therapy with a Stem Cell-Seeded Graft: Lessons from Compassionate Use Application of a GMP-Compliant Tissue-Engineered Medicine.

Tracheal replacement for the treatment of end-stage airway disease remains an elusive goal. The use of tissue-engineered tracheae in compassionate use cases suggests that such an approach is a viable option. Here, a stem cell-seeded, decellularized tissue-engineered tracheal graft was used on a compassionate basis for a girl with critical tracheal stenosis after conventional reconstructive techniques failed. The graft represents the first cell-seeded tracheal graft manufactured to full good manufacturing practice (GMP) standards. We report important preclinical and clinical data from the case, which ended in the death of the recipient. Early results were encouraging, but an acute event, hypothesized to be an intrathoracic bleed, caused sudden airway obstruction 3 weeks post-transplantation, resulting in her death. We detail the clinical events and identify areas of priority to improve future grafts. In particular, we advocate the use of stents during the first few months post-implantation. The negative outcome of this case highlights the inherent difficulties in clinical translation where preclinical in vivo models cannot replicate complex clinical scenarios that are encountered. The practical difficulties in delivering GMP grafts underscore the need to refine protocols for phase I clinical trials. Stem Cells Translational Medicine 2017;6:1458-1464.

Pediatric mechanical circulatory support: available devices and outcomes as bridge-to-transplant therapy.

PURPOSE OF REVIEW: The use of mechanical circulatory support in children has increased dramatically over the current decade. A review of the pediatric ventricular assist device (VAD) literature is timely and relevant in order to improve outcomes and refine patient selection as new devices become available and current VADs are used in smaller patients. RECENT FINDINGS: Outcomes of pediatric patients bridged to transplant with VAD therapy continue to improve. Patient selection in children continues to be complex as size and anatomy make decision-making unique from the adult practice. In addition, device selection and surgical planning are evolving in children. The use of temporary devices for long-term use is an example of an innovative application of current technology. Sensitization and adverse events are important issues that are necessary to understand in order to improve outcomes. The importance of a national registry to help monitor patient outcomes is critical to assessing this highly technological therapy. SUMMARY: There are many implications of improving and expanding durable device use in children. VAD therapy is evolving as a standard of care as a bridge to transplant in children. Continued surveillance of outcomes is critical to improving patient selection and decreasing adverse events in children with advanced heart failure.

Donante vivo. Legislación / Living donor. Legislation

Introducción: Trasplantes realizados de órganos procedentes de donante vivo pueden serlo de un riñón y parciales de hígado, intestino e incluso pulmón. Si bien varían fundamentalmente en el riesgo que supone para el donante y los resultados en la supervivencia del injerto, la implicación legal es la misma. Así, me centraré en el protocolo con mayor experiencia, menor riesgo para el donante y mejores resultados: la donación renal de vivo. El trasplante renal ha demostrado ser el mejor tratamiento de la insuficiencia renal crónica en cuanto a supervivencia, calidad de vida, menores complicaciones y mejor relación coste-beneficio frente a la diálisis. Situación actual del donante vivo: El trasplante renal procedente de donante vivo se realiza en 14 de las 17 comunidades autónomas del territorio nacional. Destacan, en el año 2013, Cataluña (165 trasplantes), Andalucía (60 trasplantes), Madrid (40 trasplantes) y País Vasco (38 trasplantes), y la evolución del plan nacional de trasplante cruzado, con un incremento muy significativo en el número de trasplantes, desde su comienzo en el año 2009 con dos trasplantes renales hasta 41 trasplantes renales procedentes de donante vivo cruzado en el año 2013. Legislación: El trasplante renal de donante vivo estaba regulado en España por la Ley de trasplantes 30/1979. Esta ley regulaba la donación en vida de un órgano si es compatible con la vida y la función del órgano o parte de él es compensada por el organismo. Especifica además que el destino del órgano será su trasplante a una persona determinada. Luego siguió el Real Decreto 2070/1999, la Ley de Autonomía del Paciente 41/2002 y finalmente el Real Decreto 1723/2012 de 28 de diciembre. En Europa, la Directiva 2010/45/UE del Parlamente Europeo y del Consejo Europeo de 7 de julio de 2010 (AU)

Introduction: Organ transplants from living donors may be a partial kidney and liver, intestine and even lung. If they vary mainly in the risk to the donor and results in graft survival, the legal implication is the same. So I will focus for this chapter in the protocol with more experience, the less risk to the donor and better results: the living kidney donation. Kidney transplantation has proven to be the best treatment of chronic renal failure in terms of survival, quality of life, fewer complications and better cost-benefit ratio compared to dialysis. Current status of the living donor: Kidney transplant from a living donor is performed in 14 of the 17 regions of the country. Highlighted in the year 2013 Catalonia (165 transplantations), Andalusia (60 transplants), Madrid (40 transplants) and the Basque Country (38 transplants). The evolution of cross-national transplantation plan, with a significant increase in the number of transplants, since its inception in 2009 with two kidney transplants, 41 kidney transplants from living donors crossed in 2013. Legislation: The living donor kidney transplantation was regulated in Spain by Law 30/1979 transplant. This law regulated living donation of an organ if it is compatible with the life and function of the body or part of it offset by the body. Further specifies that the fate of organ transplantation to be a certain person. Then he followed the Royal Decree 2070/1999, Law 41/2002 of Patient Autonomy and finally Royal Decree 1723/2012, 28 December. At the European level, Directive 2010/45/EU of the European Parliament and right of the European Council of July 7, 2010 (AU)

Producción de órganos bioartificiales / Artificial organs production

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Robotic-assisted laparoscopic ovarian tissue transplantation.

OBJECTIVE: To describe a technique for frozen-banked ovarian tissue transplantation using robotic-assisted laparoscopy. DESIGN: Case study. SETTING: Academic tertiary care center. PATIENT(S): A 38-year-old patient in remission for non-Hodgkin lymphoma, whose ovarian tissue had been frozen for 3 years. INTERVENTION(S): Robotic-assisted laparoscopic transplantation of thawed ovarian cortical tissue to the remaining ovary and peritoneum. MAIN OUTCOME MEASURE(S): Resumption of spontaneous menses, follicular development, and ovulation as demonstrated by ultrasound, and serum E(2) and P levels. RESULT(S): The patient experienced cyclic spontaneous menstruation 6 months after the transplantation. Ovulation was confirmed by ultrasound and serum E(2) and P levels at month 11 after surgery. CONCLUSION(S): Robotic-assisted laparoscopic surgery may be a good, minimally invasive alternative for the ovarian tissue transplantation procedure to restore ovarian function.

Surgical experience of refined 3-cuff technique for orthotopic small-bowel transplantation in rat: a report of 270 cases.

BACKGROUND: Establishment of an instant, reproducible, and reliable rat model of a refined 3-cuff technique for performing orthotopic intestinal transplantation is reported, and the surgical skills required to perform modified surgical procedure are discussed. METHODS: A retrospective analysis was used to study 270 rat cases subject to orthotopic intestinal transplantation (OIT) performed in our transplantation center from March 2006 to March 2008. After establishing the portal vein cuff method, a conventional hand-sewn anastomosis method combination, with porto-to-portal re-establishment by cuffed anastomosis technique, was used in group 1 (n = 140), and the modified 3-cuff anastomosis method was applied in group 2 (n = 130). Statistical comparison was made between the 2 groups. RESULTS: In group 1, 97 of 140 (69.3%) recipients survived >7 days, and 69 (49.3%) survived >30 days, whereas in group 2, respective survival was 110 of 130 (84.6%) and 86 of 130 (66.2%). Average cold ischemic times in the 2 groups were 48.5 +/- 5.1 minutes and 31 +/- 3.0 minutes, respectively. There was a significant difference between the 2 groups (P <.05). In most cases, the average volume of bleeding during recipient surgery was <1 mL using the simplified 3-cuff anastomosis technique. There was shorter graft revascularization time with the new model of sutureless microanastomosis using cuff apparatus for OIT in rats compared with the control group. The method adopted in group 2 was much easier, more stable, and more feasible than that in group 1. Sixty-three rats died in 7 days, and autopsy verified the causative factors leading to death, which are summarized in the text. The results obtained were acceptable and satisfactory. Overall, there was a comparative lower incidence of complications associated with the procedure used in group 2. CONCLUSIONS: The modified 3-cuff anastomosis technique for rat OIT models has several obvious advantages, which can be summarized as follows: vascular anastomosis is stable and simplified, and blood loss is significantly decreased; natural anatomic physiologic portal graft drainage is maintained; and intraoperative mortality and postsurgical morbidity are minimized. Furthermore, technical refinement of rat OIT models established by our research team can be carried out without a microscope and can be easily implemented in the laboratory by 1 trainee with acceptable success after a short period of training. We regard it as one of the best available orthotopic small-bowel transplantation methods in rat.

The role of alemtuzumab in facilitating maintenance immunosuppression minimization following solid organ transplantation.

National registry data indicate a trend towards the incorporation of lymphocyte depletion antibody induction therapy into immunosuppressive regimens for solid organ transplantation. Depletional induction has been shown to reduce the risk of early acute rejection, but increase the risk of immune incompetence. As such, it recently has been paired with reduced maintenance immunosuppression in an effort to curb excessive immunosuppression without sacrificing low rejection rates. Alemtuzumab is a humanized CD52-specific monoclonal antibody that has been used in the setting of maintenance immunosuppression minimization. Although not specifically indicated for organ transplantation, it is now used off-label as an induction agent in approximately 10% of transplant recipients in the United States. In general, alemtuzumab is well tolerated and substantially reduces the risk of acute rejection in the first 6 months post-transplant in non-sensitized recipients. There is little evidence to support the notion that it uniquely promotes tolerance, and growing evidence that it is ineffective in the setting of allosensitization. Alemtuzumab-treated patients clearly remain dependent on maintenance immunosuppression. Long-term outcome data will be required to determine the magnitude and type of maintenance therapy that makes best use of alemtuzumab's depletional effects.

Harvesting buccal mucosa graft under local infiltration analgesia--mitigating need for general anesthesia.

For buccal mucosal graft urethroplasty, nasal or oral endotracheal intubation anesthesia is used for harvesting the graft from the oral cavity. A technique of graft harvesting under local anesthesia using 2% lidocaine solution with adrenaline (1:200,000) is described. This method requires a cooperative patient but saves the morbidity of general anesthesia.

How to monitor renal function in pediatric solid organ transplant recipients.

The aim is to review the tools for early detection of renal dysfunction after pediatric solid organ transplantation. Currently, the most widely used marker for detection of renal dysfunction involves measurement of GFR. Inulin clearance forms the "gold standard" method for measuring GFR; however, nuclear medicine methods ((51)Cr EDTA and (99)Tc DTPA isotope clearance studies) have replaced inulin clearance. The measurement of serum creatinine has a low sensitivity for the early detection of renal damage. The Schwartz formula using patient height and serum creatinine requires center-specific constants and has limitations associated with creatinine determination. These limitations may be overcome using a cystatin C-based GFR estimation. In diabetic nephropathy, and more recently in hemolytic uremic syndrome, microalbuminuria has been established as a useful screening tool for renal damage, while its predictive value in the transplantation setting needs to be established. All transplant recipients should be screened for hypertension. Early referral for ambulatory 24-h blood pressure monitoring and involvement of pediatric nephrologists should be considered. All pediatric solid organ transplant recipients receiving CNI should be screened regularly for high blood pressure and early evidence of renal damage using either GFR scans or cystatin C-based GFR estimations.

Anesthetic considerations for multivisceral transplantation.

Multivisceral transplantation is defined as the en bloc transplantation of three or more abdominal organs. Although multivisceral transplant recipients do not yet enjoy the same survival rates as renal and liver transplant recipients, this procedure can be life saving and has shown improvements in survival rates over time. Advances in immunosuppression hold promise for the future of multivisceral transplantation.