A protocol for rat kidney normothermic machine perfusion and subsequent transplantation

Czogalla, Jan; Grahammer, Florian; Puelles, Victor G.; Huber, Tobias B.

doi:10.1111/aor.13799

Cited by 4 publications

(5 citation statements)

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“…We found the optimal time period for analysis of the ER stress response in KSI was 30 min. This is a shorter time than used in rodent NMP models (1 hour in mice, 2 hours in rats) ( Czogalla et al, 2021 ). Most probably, adaptions and molecular processes occurring during NMP are missed during this shorter time.…”

Section: Discussionmentioning

confidence: 99%

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A high-throughput drug discovery pipeline to optimize kidney normothermic machine perfusion

et al. 2022

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Kidney transplantation is the only definitive therapy for end-stage kidney disease. The shortage of organs for transplantation is the main limitation of this life-saving treatment. Normothermic machine perfusion (NMP) is a novel preservation technique with the potential to increase the number of transplantable kidneys through reducing delayed graft function and organ evaluation under physiological conditions. To date, the cellular effects and possible pharmacological interventions during machine perfusion are incompletely understood. A major limitation is the technically complex, time-consuming, and small-scale replication of NMP in rodent models. To overcome this, we developed a 3D-printed, high throughput ex-vivo mouse kidney slice incubator (KSI) mimicking mouse kidney NMP by working under closely resembling conditions. KSI significantly reduced the time per experiment and increased the sample throughput (theoretical: 54 incubations with n = 500/day). The model recapitulated the cellular responses during NMP, namely increased endoplasmic reticulum stress (ER stress). Using KSI, five pharmacological interventions against ER stress taken from the literature were tested. While four were ineffective and excluded, one, β-Nicotinamide-adenine-dinucleotide (NADH), ameliorated ER stress significantly during KSI. The test of NADH in mouse kidney NMP replicated the positive effects against ER stress. This suggests that testing the addition of NADH during clinical kidney NMP might be warranted.

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Section: Discussionmentioning

confidence: 99%

“…We have recently established a cell-free rodent NMP model for the rigorous preclinical study of potential interventions during NMP ( Czogalla et al, 2021 ). However, the complexity and time-consuming nature of rodent NMP models only allows small-scale sample generation, resulting in slow progress.…”

Section: Introductionmentioning

confidence: 99%

A high-throughput drug discovery pipeline to optimize kidney normothermic machine perfusion

et al. 2022

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“…An autologous kidney transplant model in rats was first described and established in this study (Figure 1). Jan et al 19 showed a novel kidney normothermic machine perfusion model, and the abdominal aorta was also the entry point for perfusion. The operation of the present study is not complex, with a high success rate of modeling.…”

Section: Discussionmentioning

confidence: 99%

Protective effect of estradiol copreservation against kidney ischemia‐reperfusion injury

Zhou

Leung

et al. 2021

Artificial Organs

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Ischemia-reperfusion injury (IRI) is the major cause of delayed graft function (DGF) during the posttransplantation period. Estradiol (E2) prevents IRI-induced kidney dysfunction and tissue injury. However, many side effects limit E2's in vivo application. Recent evidence uncovers E2's expanded use in the field of transplantation.We aimed to study if and how E2 exerts protective activity during the period of kidney organ preservation. The autologous kidney transplant model in rats was first established. Rats were divided into 5 groups: normal group (N), sham group (sham), static cold storage (SCS) 4 hours group (control), SCS 4 hours + ethanol (1 µL/ mL) group (solvent), and SCS 4 hours + ethanol (1 µL/mL) + E2 (1000 ng/mL) group (E2). ERα expression under hypothermia was measured by western blotting.Moreover, biochemical analyses of plasma levels of creatinine, BUN, estradiol, and testosterone were examined. Among all groups, kidney tissues were collected and processed for further western blot analysis about ERα, eNOS, Bcl-2, and Bax expression, histological analyses such as H&E staining to evaluate pathological severity.In addition, a TUNEL assay is performed to evaluate apoptosis. E2 copreservation upregulated ERα expression under hypothermia. Moreover, E2 copreservation reduced levels of creatinine and BUN in plasma but without affecting estradiol and testosterone. Further, E2 copreservation increased expression of eNOS and antiapoptotic Bcl-2 and decreases expression of proapoptotic Bax. E2 copreservation significantly inhibited IRI-induced apoptosis and evidently improved pathological severity in the kidney of rats. E2 copreservation exerts protective activity against IRI-induced pro-inflammatory and proapoptotic effects in kidneys during organ preservation time and improves transplanted kidney function.

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“…En general, esta técnica permite la evaluación continua de la tasa de flujo renal, la resistencia vascular, la liberación de hormonas, el análisis de gases en sangre, el análisis de orina y la aplicación de fármacos. Después del procedimiento, el riñón puede ser procesado para biología molecular, estudios metabolómicos, análisis bioquímico o microscopía [5].…”

Section: Introductionunclassified

Estandarización de modelo de perfusión renal en ratones

Yángüez

Morán

Mero

et al. 2021

apanac

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La técnica de riñón ex vivo permite valorar la respuesta renal intrínseca permitiendo que las variables experimentales puedan ser controladas por el investigador. En los laboratorios del CIPFAR trabajamos para poner a punto el modelo de riñón aislado y perfundido, para lo cual empleamos ratones C57BL/6 y CD-1 machos. Realizamos laparatomía abdominal, localizamos y ligamos la sección aórtica, seguidamente, realizamos una incisión en la arteria renal izquierda e introdujimos una cánula acoplada a un microtubo de polietileno. A través de la cánula, los riñones aislados son conectados a un sistema de perfusión y expuestos a los contracturantes KCl (80mM) y fenilefrina (FE 10-5 M). También obtuvimos curvas concentración respuesta frente a acetilcolina (ACh 10-8–10-4M). Nuestros resultados revelan valores de presión basal de 36.8 ± 4.8 mmHg y 30.7 ± 14.2 mmHg para las cepas CD-1 y C57BL/6, respectivamente. La despolarización con KCl desarrolló un efecto contracturante máximo de 25.7 ± 16.0 mmHg para CD-1 y 63.8 ± 34.8 mmHg en ratones C57BL/6. El agente adrenérgico FE, produjo contracciones máximas de 53.0 ± 29.6 mmHg en CD-1 y 66.1 ± 4.7 mmHg en C57BL/6. El efecto vasodilatador máximo inducido por ACh fue de 35.7 % ± 3.5 y 46.9 % ± 18.7 en CD-1 y C57BL/6, respectivamente.A la luz de este resultado, tanto el tiempo de exposición, la manipulación excesiva, la perfusión inadvertida de burbujas y la formación de coágulos, pueden ser factores que disminuyan la viabilidad del endotelio renal. En cualquier caso, los datos de perfusión basal, así como la reactividad frente a agentes contracturantes son pruebas de la viabilidad del tejido en ambos grupos de animales.

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A protocol for rat kidney normothermic machine perfusion and subsequent transplantation

Cited by 4 publications

References 35 publications

A high-throughput drug discovery pipeline to optimize kidney normothermic machine perfusion

A high-throughput drug discovery pipeline to optimize kidney normothermic machine perfusion

Protective effect of estradiol copreservation against kidney ischemia‐reperfusion injury

Estandarización de modelo de perfusión renal en ratones

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