Lennart Wiman (photographer) is acknowledged for excellent video recording.

Pazienti hanno fornito consenso informato scritto e lo studio è stato approvato dalla Regione Ethical Review Board presso l&#39;Università di Göteborg. 1. Anestesia <ol><li> Impostare tutto il monitoraggio (ECG, pressione sanguigna bracciale) 17. </li><li> Inserire un catetere epidurale in basso a metà livello toracico 17. </li><li> Indurre l&#39;anestesia con propofol 2 mg / kg, 150 mg di fentanyl, e Rocuronium 0,6 mg / kg attraverso un catetere venoso periferico precedentemente inserito. </li><li> Preoxygenate e intubare il paziente. Mantenere l&#39;anestesia con sevoflurano, l&#39;ossigeno e l&#39;aria 17. </li><li> Inserire un doppio lume catetere venoso centrale e un catetere arterioso. Monitorare CVP e pressione arteriosa 17. </li></ol> 2. Isolamento del Fegato e Incannulazione delle navi <ol><li> Somministrare 100 IE / kg di eparina e dare profilassi antibiotica (ad esempio, 160 mg di trimetoprim e 800 mg di Sulfamethoxazole). </li><li> Utilizzando la tecnica Seldinger percutanea, posizionare un catetere P. 17 nella vena femorale e un padre del catetere 15 nella vena giugulare esterna 18. Verificare le posizioni dei cateteri dalla fluoroscopia. </li><li> Eseguire una laparotomia da un&#39;incisione a forma di L dal processo xifoideo lungo la linea mediana e poi esteso a un diritto un&#39;incisione sottocostale 19. </li><li> Verificare che non vi siano metastasi extra epatica nell&#39;addome per una completa ispezione e palpazione. <ol><li> Se vengono rilevati lesioni sospette, verificare malignità da sezioni congelate. </li><li> Terminare la procedura se vi è evidenza di manifestazione extraepatica (adenopatia periaortal, metastasi addominali). Linfoadenopatia nel epatico non è considerata una controindicazione. </li></ol></li><li> Mobilitare il fegato dividendo il legamento falciforme, il legamento triangolare di sinistra e destra e il legamento coronario 19. </li><li> Sezionare il Cavalvenosa partire dalle vene renali fino al diaframma con legatura delle vene tributarie retroperitoneali. </li><li> Identificare il diritto vena surrenalica e mettere un cappio nave intorno ad esso. </li><li> Identificare il diritto della vena gonadica. Sezionarlo per 5-10 cm. Collocare una sutura a borsa di intorno all&#39;ingresso della vena cava. <ol><li> Posizionare una legatura distale sulla vena gonadica. Utilizzando forbici, aprire la vena gonadica 3-5 cm dalla vena cava. </li><li> Inserire un 18 Fr catetere filo-rinforzato attraverso il diritto della vena gonadica e posizionare la punta retrohepatically sopra le vene renali. </li><li> Inserire una legatura intorno alla vena gonadica per fissare la posizione del catetere. </li><li> Verifica ritorno del sangue attraverso il catetere. Eparinizzare il catetere 18. </li></ol></li><li> Sezionare il legamento epatoduodenale e visualizzare l&#39;arteria epatica comune, l&#39;arteria gastroduodenale e l&#39;arteria epatica propria. <ol><li> Luogo nave loop intorno arte epatico comunery e l&#39;arteria gastrodoudenal. </li><li> Sezionare la vena porta e mettere un cappio nave intorno ad esso. </li><li> Isolare il resto del legamento contenente il dotto biliare comune e comprendono tutto questo in un ciclo vaso. </li></ol></li><li> Sollevare l&#39;arteria gastroduodenale con il ciclo nave. Dopo l&#39;interruzione del flusso arterioso, fare un&#39;incisione con le forbici. <ol><li> Inserire un 8-12 Fr catetere con la sua punta nell&#39;arteria epatica propria. Fissare la posizione con una legatura intorno all&#39;arteria gastroduodenale. </li><li> Verificare ritorno del sangue nel catetere e poi eparinizzare esso 18. </li></ol></li><li> Prime il sistema di perfusione con una unità di eritrociti confezionati, 100 ml di albumina (50 g / L), 100 ml di Tribonat (bicarbonato di sodio), 2500 unità di eparina (5.000 IE / ml) e 250 ml di cristalloidi. </li><li> Collegare arterioso e cateteri vena cava al sistema di perfusione costituiti da una macchina cuore-polmone dotato di una pompa a rulli e un riscaldatore together con venose sistemi reservoir, ossigenatori e tubi di tipo usa e getta. </li><li> Collegare i cateteri femorali giugulari ed esterni alla pompa bypass venovenosa costituito da una pompa centrifuga, con un riscaldatore e iniziare bypass. </li><li> Completare l&#39;isolamento del fegato stringendo un laccio emostatico Rumel intorno alla vena cava infrahepatically sopra le vene renali e bloccare la vena cava suprahepatically utilizzando un morsetto vascolare Klintmalm. <ol><li> Stringere un laccio emostatico Rumel intorno al dotto biliare comune, la vena porta e la vena surrenalica destra e infine bloccare l&#39;arteria epatica con un morsetto vascolare. </li></ol></li></ol> 3. La perfusione <ol><li> Avviare la perfusione e controllare manualmente il livello ematico nel serbatoio venoso. Regolare il flusso fino a raggiungere un livello di stato stazionario, di solito una portata tra 500-1,200 ml / min. </li><li> Mettere sonde termistore nei giusti e fegato lasciato lobi e uno direttamente inil catetere arterioso. </li><li> Riscaldare il perfusato mira a una temperatura obiettivo di fegato di 40 o C. Mantenere questa temperatura per tutta la perfusione. </li><li> Posizionare una sonda scintillazione sopra la testa della pompa bypass venovenosa. </li><li> Calibrare il sistema di monitoraggio di dispersione, iniettando una piccola dose di 10 MBq 99m Tc-albumina (1 ml) nella circolazione sistemica e registrare l&#39;attività basale. <ol><li> Iniettare un dieci volte più alta dose, 100 MBq 99m Tc-albumina (1 ml), nell&#39;unità di perfusione epatica. </li><li> Registra qualsiasi fuoriuscita dal sistema di perfusione alla circolazione sistemica come un aumento dell&#39;attività misurata dalla sonda scintillazione. </li><li> Registra qualsiasi perdita dalla circolazione sistemica al sistema di perfusione come un aumento del volume nel serbatoio venoso. </li></ol></li><li> Quando viene stabilita stato stazionario della portata, nessuna perdita è presente e la temperatura del fegato ha raggiunto 40 ° C, iniettare melphalan alla dose di1 mg / kg di peso corporeo nel sistema di perfusione diviso in due dosi somministrate 30 min a parte. </li></ol> 4. Terminare la perfusione e Rimuovi Cateteri <ol><li> Dopo 60 min di perfusione, svuotare il perfusato dal sistema e irrigare il fegato con 1.000 ml di crystalloid. </li><li> Aggiungere una unità di eritrociti confezionati all&#39;unità perfusione, infondere nel fegato e quindi fermare la perfusione. </li><li> Rilasciare il morsetto Klintmalm e il laccio emostatico Rumel dalla vena cava. </li><li> Rilasciare il laccio emostatico Rumel dal legamento epatoduodenale. </li><li> Rimuovere la legatura dalla arteria gastroduodenale. Rimuovere il catetere e legare il vaso. </li><li> Rilasciare il morsetto dalla arteria epatica. </li><li> Rimuovere il catetere dalla vena gonadica e legare il vaso. </li><li> Fermare la pompa di bypass veno-venosa e scollegare i tubi. </li><li> Chiudere l&#39;addome con una sutura fasciale ininterrotta e poi una sutura cutanea intracutaneous. </li><li> Rimuovere la catheters nelle vene giugulari femorali ed esterni e applicare pressione locale per 5 min. </li></ol> 5. Cura postoperatoria <ol><li> Estubare il paziente e il trasferimento di unità di terapia post-operatoria per 24 ore. Mantenere il controllo della pressione venosa centrale e pressione arteriosa. </li><li> Trasferire il paziente in un reparto di chirurgia per un tipico di 3-7 giorni di cure post-operatorie. </li><li> Continuare con basso peso molecolare eparina (LMWH) alla dose di 5000 IE / giorno per un totale di 4 settimane. </li></ol>

<ol>
	<li>Virgili, G., 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=Incidence+of+uveal+melanoma+in+Europe.">Incidence of uveal melanoma in Europe.</a> Ophthalmology. 114 (12), 2309-2315 (2007).</li><li>Pereira, P. 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=Current+and+emerging+treatment+options+for+uveal+melanoma.">Current and emerging treatment options for uveal melanoma.</a> Clinical Ophthalmology. 7, 1669-1682 (2013).</li><li>Bergman, L., Nilsson, B., Lundell, G., Lundell, M., Seregard, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ruthenium+brachytherapy+for+uveal+melanoma,+1979-2003:+survival+and+functional+outcomes+in+the+Swedish+population.">Ruthenium brachytherapy for uveal melanoma, 1979-2003: survival and functional outcomes in the Swedish population.</a> Ophthalmology. 112 (5), 834-840 (2005).</li><li>Diener-West, M., 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=Development+of+metastatic+disease+after+enrollment+in+the+COMS+trials+for+treatment+of+choroidal+melanoma:+Collaborative+Ocular+Melanoma+Study+Group+Report+No.+26.">Development of metastatic disease after enrollment in the COMS trials for treatment of choroidal melanoma: Collaborative Ocular Melanoma Study Group Report No. 26.</a> Archives of Ophthalmology. 123 (12), 1639-1643 (2005).</li><li>Kujala, E., Makitie, T., Kivela, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Very+long-term+prognosis+of+patients+with+malignant+uveal+melanoma.">Very long-term prognosis of patients with malignant uveal melanoma.</a> Investigative Ophthalmology &#38; Visual Science. 44 (11), 4651-4659 (2003).</li><li>Bedikian, A. Y., Papadopoulos, N., Plager, C., Eton, O., Ring, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+II+evaluation+of+temozolomide+in+metastatic+choroidal+melanoma.">Phase II evaluation of temozolomide in metastatic choroidal melanoma.</a> Melanoma Research. 13 (3), 303-306 (2003).</li><li>Kivela, T., 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=vincristine,+lomustine+and+dacarbazine+(BOLD)+in+combination+with+recombinant+interferon+alpha-2b+for+metastatic+uveal+melanoma.">vincristine, lomustine and dacarbazine (BOLD) in combination with recombinant interferon alpha-2b for metastatic uveal melanoma.</a> European Journal Of Cancer. 39 (8), 1115-1120 (2003).</li><li>Schmittel, A., 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=Phase+II+trial+of+cisplatin,+gemcitabine+and+treosulfan+in+patients+with+metastatic+uveal+melanoma.">Phase II trial of cisplatin, gemcitabine and treosulfan in patients with metastatic uveal melanoma.</a> Melanoma Research. 15 (3), 205-207 (2005).</li><li>Leyvraz, S., 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=Hepatic+intra-arterial+versus+intravenous+fotemustine+in+patients+with+liver+metastases+from+uveal+melanoma+(EORTC+18021):+a+multicentric+randomized+trial.">Hepatic intra-arterial versus intravenous fotemustine in patients with liver metastases from uveal melanoma (EORTC 18021): a multicentric randomized trial.</a> Annals Of Oncology : Official Journal Of The European Society for Medical Oncology / ESMO. 25 (3), 742-746 (2014).</li><li>Huppert, P. E., 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=Transarterial+chemoembolization+of+liver+metastases+in+patients+with+uveal+melanoma.">Transarterial chemoembolization of liver metastases in patients with uveal melanoma.</a> European Journal Of Radiology. 74 (3), 38-44 (2010).</li><li>Klingenstein, A., Haug, A. R., Zech, C. J., Schaller, U. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radioembolization+as+locoregional+therapy+of+hepatic+metastases+in+uveal+melanoma+patients.">Radioembolization as locoregional therapy of hepatic metastases in uveal melanoma patients.</a> Cardiovascular and Interventional Radiology. 36 (1), 158-165 (2013).</li><li>Mariani, P., 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=Surgical+management+of+liver+metastases+from+uveal+melanoma:+16+years'+experience+at+the+Institut+Curie.">Surgical management of liver metastases from uveal melanoma: 16 years' experience at the Institut Curie.</a> European Journal Of Surgical Oncology : The Journal Of The European Society Of Surgical Oncology And The British Association of Surgical Oncology. 35 (11), 1192-1197 (2009).</li><li>Ausman, R. K., Aust, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolated+perfusion+of+the+liver+with+HN2.">Isolated perfusion of the liver with HN2.</a> Surgical Forum. 10, 77-79 (1960).</li><li>Dahl, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+of+hyperthermia+and+chemotherapy.">Interaction of hyperthermia and chemotherapy.</a> Recent results in cancer research. Fortschritte der Krebsforschung. Progres Dans Les Recherches Sur Le Cancer. 107, 157-169 (1988).</li><li>Barker, W. C., Andrich, M. P., Alexander, H. R., Fraker, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuous+intraoperative+external+monitoring+of+perfusate+leak+using+iodine-131+human+serum+albumin+during+isolated+perfusion+of+the+liver+and+limbs.">Continuous intraoperative external monitoring of perfusate leak using iodine-131 human serum albumin during isolated perfusion of the liver and limbs.</a> European Journal Of Nuclear Medicine. 22 (11), 1242-1248 (1995).</li><li>Olofsson, 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=Isolated+hepatic+perfusion+for+ocular+melanoma+metastasis:+registry+data+suggests+a+survival+benefit.">Isolated hepatic perfusion for ocular melanoma metastasis: registry data suggests a survival benefit.</a> Annals of Surgical Oncology. 21 (2), 466-472 (2014).</li><li>Redai, I., Emond, J., Brentjens, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthetic+considerations+during+liver+surgery.">Anesthetic considerations during liver surgery.</a> The Surgical Clinics of North America. 84, 401-411 (2004).</li><li>Sakai, T., 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=Insertion+and+management+of+percutaneous+veno-venous+bypass+cannula+for+liver+transplantation:+a+reference+for+transplant+anesthesiologists.">Insertion and management of percutaneous veno-venous bypass cannula for liver transplantation: a reference for transplant anesthesiologists.</a> Clinical Transplantation. 24 (5), 585-591 (2010).</li><li>Skandalakis, J. E., Skandalakis, L. J., Skandalakis, P. N., Mirilas, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+surgical+anatomy.">Hepatic surgical anatomy.</a> The Surgical Clinics of North America. 84 (2), 413-435 (1016).</li><li>Alexander, H. 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=A+phase+I-II+study+of+isolated+hepatic+perfusion+using+melphalan+with+or+without+tumor+necrosis+factor+for+patients+with+ocular+melanoma+metastatic+to+liver.">A phase I-II study of isolated hepatic perfusion using melphalan with or without tumor necrosis factor for patients with ocular melanoma metastatic to liver.</a> Clinical Cancer Research : An Official Journal Of The American Association for Cancer Research. 6 (8), 3062-3070 (2000).</li><li>Alexander, H. R., Jr, , 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=Hyperthermic+isolated+hepatic+perfusion+using+melphalan+for+patients+with+ocular+melanoma+metastatic+to+liver.">Hyperthermic isolated hepatic perfusion using melphalan for patients with ocular melanoma metastatic to liver.</a> Clinical Cancer Research : An Official Journal Of The American Association For Cancer Research. 9 (17), 6343-6349 (2003).</li><li>Rizell, M., 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=Isolated+hepatic+perfusion+for+liver+metastases+of+malignant+melanoma.">Isolated hepatic perfusion for liver metastases of malignant melanoma.</a> Melanoma research. 18 (2), 120-126 (2008).</li><li>Iersel, L. 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=Isolated+hepatic+perfusion+with+200+mg+melphalan+for+advanced+noncolorectal+liver+metastases.">Isolated hepatic perfusion with 200 mg melphalan for advanced noncolorectal liver metastases.</a> Annals of Surgical Oncology. 15 (7), 1891-1898 (2008).</li><li>Carvajal, R. D., 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=Effect+of+selumetinib+vs+chemotherapy+on+progression-free+survival+in+uveal+melanoma:+a+randomized+clinical+trial.">Effect of selumetinib vs chemotherapy on progression-free survival in uveal melanoma: a randomized clinical trial.</a> JAMA : the Journal Of The American Medical Association. 311 (23), 2397-2405 (2014).</li><li>Sharma, K. V., 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=Hepatic+arterial+chemoembolization+for+management+of+metastatic+melanoma.">Hepatic arterial chemoembolization for management of metastatic melanoma.</a> AJR. American Journal Of Roentgenology. 190 (1), 99-104 (2008).</li><li>Eldredge-Hindy, H., 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=Yttrium-90+Microsphere+Brachytherapy+for+Liver+Metastases+From+Uveal+Melanoma:+Clinical+Outcomes+and+the+Predictive+Value+of+Fluorodeoxyglucose+Positron+Emission+Tomography.">Yttrium-90 Microsphere Brachytherapy for Liver Metastases From Uveal Melanoma: Clinical Outcomes and the Predictive Value of Fluorodeoxyglucose Positron Emission Tomography.</a> American Journal Of Clinical Oncology. , (2014).</li><li>Etten, 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=Isolated+hypoxic+hepatic+perfusion+with+orthograde+or+retrograde+flow+in+patients+with+irresectable+liver+metastases+using+percutaneous+balloon+catheter+techniques:+a+phase+I+and+II+study.">Isolated hypoxic hepatic perfusion with orthograde or retrograde flow in patients with irresectable liver metastases using percutaneous balloon catheter techniques: a phase I and II study.</a> Annals of Surgical Oncology. 11 (6), 598-605 (2004).</li><li>Ku, Y., 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=Extracorporeal+adriamycin-removal+following+hepatic+artery+infusion:+use+of+direct+hemoperfusion+combined+with+veno-venous+bypass.">Extracorporeal adriamycin-removal following hepatic artery infusion: use of direct hemoperfusion combined with veno-venous bypass.</a> Nihon Geka Gakkai Zasshi. 90 (10), 1758-1764 (1989).</li><li>Curley, S. A., 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=Hepatic+arterial+infusion+chemotherapy+with+complete+hepatic+venous+isolation+and+extracorporeal+chemofiltration:+a+feasibility+study+of+a+novel+system.">Hepatic arterial infusion chemotherapy with complete hepatic venous isolation and extracorporeal chemofiltration: a feasibility study of a novel system.</a> Anti-Cancer Drugs. 2 (2), 175-183 (1991).</li><li>Beheshti, M. V., 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=Percutaneous+isolated+liver+perfusion+for+treatment+of+hepatic+malignancy:+preliminary+report.">Percutaneous isolated liver perfusion for treatment of hepatic malignancy: preliminary report.</a> Journal of Vascular And Interventional Radiology : JVIR. 3 (3), 453-458 (1992).</li></ol>

The authors have nothing to disclose.

I pazienti con melanoma metastatico uveale hanno poche opzioni terapeutiche. Chemioterapia 6, immunoterapia 23 e terapia mirata 24 non sono stati in grado di mostrare una migliore OS. Per i pazienti con poche metastasi epatiche, resezione epatica è un&#39;opzione 12. Altri trattamenti includono la chemioterapia intra-arteriosa epatica (HIA) 9, chemioembolizzazione transarteriosa (TACE) 25 e la terapia selettiva interna di radiazioni (SIRT) 26....

Il melanoma uveale è il tumore maligno intraoculare primario più comune negli adulti, con l&#39;essere la più alta incidenza nelle popolazioni caucasiche 1. Trattamento locale si compone di enucleazione o placca locale radioterapia 2,3. Indipendentemente dal trattamento locale, metastasi in definitiva in circa il 35-50% dei pazienti 4,5. Il fegato è il sito più comune di metastasi e circa il 50% dei pazienti presentano isolate metastasi epatiche. La sopravvivenza globale mediana (OS) per i pazienti con metastasi epatiche è di 6-12 mesi 4. Diversi diverse strategie di trattamento sistemici e regionali sono state esplorate, ma i tassi di sopravvivenza non sono migliorati 2. Il trattamento con la chemioterapia sistemica, sia singola sostanza (dacarbazina o temozolomide 6) o trattamenti di combinazione (regime BOLD 7, gemcitabina con Treosulfan 8), mostra tassi di risposta inferiori poi10%. Epatica chemioterapia intra-arteriosa (HIA) ha mostrato un tasso leggermente superiore complessiva risposta rispetto alla chemioterapia sistemica, ma nessun aumento della sopravvivenza globale 9. Chemioembolizzazione transarteriosa (TACE) con cisplatino e carboplatino ha mostrato una risposta parziale nel 57% dei pazienti con una sopravvivenza mediana di 9 mesi 10. Radioterapia interna selettiva (SIRT) ha mostrato una risposta parziale nel 62% dei pazienti con una sopravvivenza mediana di 7 mesi 11. In uno studio su resezione epatica, inclusi 255 pazienti, è stata riportata una sopravvivenza complessiva di 14 mesi; in 76 dei pazienti in cui una resezione microscopicamente completo è stato possibile, la sopravvivenza era 27 mesi 12. Perfusione epatica isolata (IHP) è una opzione di trattamento regionale, originariamente sviluppato da Ryan e Ausman. La logica alla base della tecnica è quello di isolare chirurgicamente e profumato fegato con una dose elevata di un agente chemioterapico, evitando sistemicoxicity 13. Inoltre, ipertermia concomitante media un maggiore assorbimento dell&#39;agente chemioterapico da variazioni del flusso sanguigno tumorale e la permeabilità delle membrane cellulari 14. Completo isolamento vascolare è confermata con una tecnica di tracciante radioattivo con Tc-99 etichettata albumina sierica umana 15. Una retrospettiva studio di fase II ha mostrato un potenziale beneficio di sopravvivenza di 14 mesi (26 vs 12 mesi) per i pazienti sottoposti IHP rispetto ai sopravvissuti più lunghe con metastasi epatiche melanoma uveale in Svezia 16. Ecco a voi la tecnica di IHP con una breve rassegna dei risultati attuali e prospettive future.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Wire re-inforced catethers</td>
			<td>Medtronic Inc, Minnesota, USA</td>
			<td></td>
			<td>12-18Fr</td>
		</tr>
		<tr>
			<td>St&#246;ckert S5</td>
			<td>Sorin Group, Mirandola, Italy</td>
			<td></td>
			<td>Heart-lung machine</td>
		</tr>
		<tr>
			<td>Rotaflow</td>
			<td>Maquet, Jostra Medizintechnik AG, Hirrlingen, Germany</td>
			<td></td>
			<td>Centrifugal pump</td>
		</tr>
		<tr>
			<td>HU 35</td>
			<td>Maquet cardiopulmonary AG, Hirrlingen, Germany</td>
			<td></td>
			<td>Heater unit</td>
		</tr>
		<tr>
			<td>Flexbumin 200 g/l</td>
			<td>Baxter Medical AB, Kista, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tribonat</td>
			<td>Fresenius Kabi AB, Uppsala, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin LEO, 5000 IE/ml</td>
			<td>LEO Pharma AB, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer Acetate</td>
			<td>Baxter Medical AB, Kista, Sweden</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vasculosis</td>
			<td>Cis-Bio International, Gif-Sur-Yvette, France</td>
			<td></td>
			<td>99mTc-albumin</td>
		</tr>
		<tr>
			<td>MedicView</td>
			<td>SystemData AB, Gothenburg, Sweden</td>
			<td></td>
			<td>Leakage monitoring system</td>
		</tr>
		<tr>
			<td>Alkeran</td>
			<td>Aspen Europe GmbH, Bad Oldesloe, Germany</td>
			<td></td>
			<td>melphalan</td>
		</tr>
	</tbody>
</table>

isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma

Isolato perfusione epatica (IHP) è una procedura in cui il fegato è chirurgicamente isolato e perfuso con un&#39;alta concentrazione di melfalan agente chemioterapico. In breve, la procedura inizia con la messa a punto di un veno-venosa bypass percutanea dalla vena femorale alla vena giugulare esterna. Via una laparotomia, cateteri vengono poi inseriti in arteria epatica propria e la vena cava. La vena porta e la vena cava, sia sopra- e infrahepatically, vengono poi bloccato. Il arteriosa e cateteri venosi sono collegati ad una macchina cuore-polmone e fegato vengono perfuse con melfalan (1 mg / kg di peso corporeo) per 60 min. In questo modo è possibile defluire in localmente fegato con una dose elevata di un agente chemioterapico, senza perdite nella circolazione sistemica. In studi precedenti, inclusi i pazienti con metastasi epatiche isolate di melanoma uveale, un tasso di risposta complessiva del 33-100% e una sopravvivenza media tra i 9 ei 13 mesi,sono stati riportati. L&#39;obiettivo di questo protocollo è quello di fornire una chiara descrizione di come eseguire la procedura e discutere IHP come opzione di trattamento per le metastasi epatiche di melanoma uveale.

Una prima relazione comprendeva 22 pazienti trattati con melfalan con o senza TNF-alfa. I risultati hanno mostrato un tasso di risposta globale del 62%, tra cui due pazienti con CR (10%), e una sopravvivenza globale mediana di 11 mesi 20. Uno studio successivo ha riferito sui risultati di 29 pazienti consecutivi trattati con melfalan solo. Il risultato è stato molto simile, con una risposta globale del 62% di cui 10% di risposta completa e una sopravvivenza media di 12 mesi 21. <p class="jove_...

Watch this Scientific Journal Video about Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma at JoVE.com

Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

Isolata epatica perfusione come Trattamento di metastasi epatiche di melanoma uveale

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

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13.3K Views.  Institute of Clinical Sciences. Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

Video: Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Procedure chirurgiche per un modello di Rat parziale trapianto di fegato con la ricostruzione arteriosa epatica

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Ricerca

Medicina

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

Un preclinici Modello murino di metastasi epatiche

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

MRI-guided dmPFC-rTMS as a Treatment for Treatment-resistant Major Depressive Disorder

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial prefrontal cortex (dmPFC) in patients with major depressive disorder (MDD). Technicians used a neuronavigation system to process patient MRIs to generate a 3-dimensional head model. The head model was subsequently used to identify patient-specific stimulatory targets. The dmPFC was stimulated daily for 20 sessions. Stimulation intensity was titrated to address scalp pain associated with rTMS. Weekly assessments were conducted on the patients using the Hamilton Rating Scale for Depression (HamD17) and Beck Depression Index II (BDI-II). Treatment-resistant MDD patients achieved significant improvements on both HAMD and BDI-II. Of note, angled, double-cone coil rTMS at 120% resting motor threshold allows for optimal stimulation of deeper midline prefrontal regions, which results in a possible therapeutic application for MDD. One major limitation of the rTMS field is the heterogeneity of treatment parameters across studies, including duty cycle, number of pulses per session and intensity. Further work should be done to clarify the effect of stimulation parameters on outcome. Future dmPFC-rTMS work should include sham-controlled studies to confirm its clinical efficacy in MDD.

Here we outline the procedure for MRI-guided repetitive transcranial magnetic stimulation to the dorsomedial prefrontal cortex as an experimental treatment for major depressive disorder.

MRI-guidata dmPFC-rTMS come Trattamento di trattamento resistente Disturbo Depressivo Maggiore

Here we outline the protocol for magnetic resonance imaging (MRI) guided repetitive transcranial magnetic stimulation (rTMS) to the dorsal medial ...

Percutaneous Hepatic Perfusion (PHP) with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects. Previously, this was performed during open surgery with satisfying results, but morbidity and mortality related to the open procedure, prohibited a widespread application. Therefore, percutaneous hepatic perfusion (PHP) with simultaneous chemofiltration was developed. Besides decreasing morbidity and mortality, this procedure can be repeated, hopefully leading to a higher response rate and improved survival (by local control of disease). During PHP, catheters are placed in the proper hepatic artery, to infuse the chemotherapeutic agent, and in the inferior caval vein to aspirate the chemosaturated blood returning through the hepatic veins. The caval vein catheter is a double balloon catheter that prohibits leakage into the systemic circulation. The blood returning from the hepatic veins is aspirated through the catheter fenestrations and then perfused through an extra-corporeal filtration system. After filtration, the blood is returned to the patient by a third catheter in the right internal jugular vein. During PHP a high dose of melphalan is infused into the liver, which is toxic and would lead to life threatening complications when administered systemically. Because of the significant hemodynamic instability resulting from the combination of caval vein occlusion and chemofiltration, hemodynamic monitoring and hemodynamic support is of paramount importance during this complex procedure.

Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

In this manuscript, we describe percutaneous isolated hepatic perfusion with simultaneous chemofiltration as treatment for unresectable liver metastases. This procedure is performed under general anaesthesia in the angiosuite by an experienced team, consisting of an interventional radiologist, a clinical perfusionist and anaesthesiologist.

Percutanea epatica Perfusione (PHP) con melfalan come trattamento per non resecabili metastasi confinata al fegato

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn ...

Generation of a Humanized Mouse Liver Using Human Hepatic Stem Cells

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in which the liver has been repopulated with functional human hepatocytes, could serve as a useful tool for investigating human hepatic cell biology, drug metabolism, and other preclinical applications. One of the key factors required for successful transplantation of human hepatocytes into mice is the elimination of the endogenous hepatocytes to prevent competition with the human cells and provide a suitable space and microenvironment for promoting human donor cell expansion and differentiation. To date, two major liver injury mouse models utilizing fumarylacetoacetate hydrolase (Fah) and uroplasminogen activator (uPA) mice have been established. However, Fah mice are used mainly with mature hepatocytes and the application of the uPA model is limited by decreased breeding. To overcome these limitations, Alb-toxin receptor mediated cell knockout (TRECK)/SCID mice were used for in vivo differentiation of immature human hepatocytes and humanized liver generation. Human hepatic stem cells (HpSCs) successfully repopulated the livers of Alb-TRECK/SCID mice that had developed lethal fulminant hepatic failure following diphtheria toxin (DT) treatment. This model of a humanized liver in Alb-TRECK/SCID mice will have functional applications in studies involving drug metabolism and drug-drug interactions and will promote other in vivo and in vitro studies.

Here, we present a novel humanized mouse liver model generated in Alb-toxin receptor mediated cell knockout (TRECK)/SCID mice following the transplantation of immature and expandable human hepatic stem cells.

Generazione di un umanizzato mouse Fegato Utilizzando epatici umani cellule staminali

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Metodo di consegna diretta segmentale intra-epatico utilizzando un modello di fegato di ratto Hilar morsetto

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Treatment of Liver Metastases Using an Internal Target Volume Method for Stereotactic Body Radiotherapy

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For inoperable patients, local ablation therapies, such as stereotactic body radiotherapy (SBRT), can provide effective local tumor control with minimal toxicity. Because of its high precision and accuracy, SBRT delivers a higher radiation dose per fraction, is more effective, and targets smaller irradiation volumes than does conventional radiotherapy. In addition, steep dose gradients from target lesions to surrounding normal tissues are achieved using SBRT; thus, SBRT provides more effective tumor control and exhibits fewer side effects than conventional radiotherapy. The use of SBRT is prevalent for treating intracranial lesions (known as stereotactic radiosurgery); however, it is now also used for treating spinal and adrenal metastases. Because of advancements in image-guided assistance and respiratory motion management, several studies have investigated the use of SBRT for treating lung or liver tumors, which move as a patient breathes. The results of these studies have suggested that SBRT favorably controls tumors in the case of moving lesions.
Four-dimensional computed tomography (4D-CT) with an abdominal compressor (AC) is clinically convenient for effective respiratory motion management. Because this method is noninvasive and allows free breathing, its use reduces complications. Furthermore, patients consider this method convenient. Moreover, it is considered more efficient than other methods of respiratory motion management by physicians and therapists. The use of 4D-CT with an AC for treating pulmonary lesions has also been widely investigated, and the technique is gaining acceptance for treating hepatic lesions. However, the protocols for using 4D-CT with an AC for treating hepatic lesions are different from those used for treating pulmonary lesions. In this article, we describe a new protocol for SBRT with 4D-CT and an AC for treating liver metastases.

Stereotactic body radiotherapy (SBRT) requires rigorous accuracy and precision for delivering high radiation doses per fraction to small treatment volumes to improve tumor control and simultaneously reduce toxicity. Herein, we present a noninvasive and clinically convenient respiratory motion management protocol for SBRT for liver metastases.

Trattamento delle metastasi del fegato usando un metodo di Volume bersaglio interno per la radioterapia stereotassica corpo

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

Un piccolo modello animale di aspersione epatica normotermica Ex Vivo

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

Misurazione del gradiente di pressione venosa epatica e della biopsia epatica transgiugulare

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

Murine Precision-Cut Liver Slices come un modello Ex Vivo di biologia del fegato

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...