Name: isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma
Uploaded: 2015-01-25T14:00:00.000+00:00
Duration: 9 min 52 s
Description: Watch this Scientific Journal Video about Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma at JoVE.com

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

Patienten mit schriftlicher Einverständniserklärung und die Studie wurde von der Regional Ethical Review Board an der Universität Göteborg genehmigt. 1. Anästhesie <ol><li> Richten Sie alle Überwachung (EKG, Blutdruckmanschette) 17. </li><li> Legen Sie eine Epiduralkatheter im unteren bis mittleren thorakalen Ebene 17. </li><li> Induzieren Narkose mit Propofol 2 mg / kg, 150 &amp; mgr; g Fentanyl und Rocuronium 0,6 mg / kg durch einen zuvor einge peripheren Venenkatheter. </li><li> Preoxygenate und Intubation des Patienten. Aufrechterhaltung einer Narkose mit Sevofluran, Sauerstoff und Luft 17. </li><li> Legen Sie eine Doppellumen-ZVK und einen arteriellen Katheter. Überwachen CVP und arteriellen Druck 17. </li></ol> 2. Isolierung der Leber und Kanülierung der Gefäße <ol><li> Verwalten 100 IE / kg Heparin und geben Antibiotikaprophylaxe (zB 160 mg Trimethoprim und 800 mg Sulfamethoxazol). </li><li> Mit der perkutanen Seldinger-Technik, setzen Sie ein 17 Fr Katheter in die Vena femoralis und eine 15 Fr Katheter in der V. jugularis externa 18. Überprüfen Sie die Positionen der Katheter durch Fluoroskopie. </li><li> Durchführen einer Laparotomie durch einen L-förmigen Einschnitt von der Schwertfortsatz entlang der Mittellinie und dann auf eine richtige subcostal Einschnitt 19 erstreckt. </li><li> Stellen Sie sicher, dass keine außerLeberMetastasen vorhanden sind in den Unterleib mit einer kompletten Inspektion und Palpation. <ol><li> Wenn irgendwelche verdächtigen Läsionen gefunden werden, überprüfen Malignität durch Gefrierschnitt. </li><li> Beenden Sie das Verfahren, wenn es Anzeichen für extrahepatische Manifestation (periaortal Adenopathie, Bauch Metastasen). Lymphadenopathie in der Leberpforte wird nicht als Kontraindikation. </li></ol></li><li> Mobilisieren der Leber durch Dividieren des Ligamentum falciforme, die linken und rechten Dreiecksbandes und die koronare Ligament 19. </li><li> Präparieren Sie die Kava-Venen beginnend von den Nierenvenen bis zu der Membran mit Unterbindung der retroperitonealen Nebenvenen. </li><li> Identifizieren Sie die rechte Neben Vene und setzen Sie ein Gefäß Schleife um ihn herum. </li><li> Identifizieren Sie den rechten Gonaden Vene. Sezieren für 5-10 cm. Legen Sie eine Tabaksbeutelnaht um den Eingang zur Hohlvene. <ol><li> Legen Sie eine Ligatur distal an der Gonaden Vene. Mit einer Schere, öffnen Sie die Gonaden Vene 3-5 cm von der Hohlvene. </li><li> Legen Sie eine 18 Fr drahtverstärkte Katheter durch die rechte Gonaden Vene und legen Sie die Spitze retrohepatically über den Nierenvenen. </li><li> Legen Sie eine Ligatur um die Gonaden Vene, um die Position des Katheters zu sichern. </li><li> Überprüfen Sie den Rückfluss des Blutes durch den Katheter. Heparinisierung des Katheters 18. </li></ol></li><li> Präparieren Sie die Ligamentum hepatoduodenale und visualisieren die gemeinsame Leberarterie, der gastroduodenalen Arterie und die richtige Leberarterie. <ol><li> Platz Vesselloops um die gemeinsame Leber artery und die gastrodoudenal Arterie. </li><li> Präparieren Sie die Pfortader und setzen Sie ein Gefäß Schleife um ihn herum. </li><li> Isolieren der Rest des Bandes, die den Gallengang und umfassen all dies in einem Gefäß Schleife. </li></ol></li><li> Heben Sie den gastroduodenalen Arterie mit der Gefäßschleife. Nach Unterbrechung des arteriellen Zustroms, einen Einschnitt mit einer Schere. <ol><li> Legen Sie eine 8-12 Fr Katheter mit seiner Spitze in die richtige Leberarterie. Sichern Sie die Position mit einer Ligatur um den gastroduodenalen Arterie. </li><li> Stellen Sie sicher, Blutrück im Katheter und dann Heparinisierung es 18. </li></ol></li><li> Prime das Perfusionssystem mit einer Einheit verpackt Erythrozyten, 100 ml Albumin (50 g / l), 100 ml Tribonat (Natriumbicarbonat), 2500 Einheiten Heparin (5000 IE / ml) und 250 ml Kristalloide. </li><li> Verbinden des arteriellen und die Hohlvene Katheter zur Perfusion System bestehend aus einem Herz-Lungen-Maschine mit einer Walzenpumpe ausgestattet und einen Heizer temeinsam mit venösen Reservoir, Oxygenator und Schlauchsysteme von Einweg-Typ. </li><li> Verbinden die femoralen und externen jugularen Katheter zur venovenöse Bypasspumpe, bestehend aus einer Zentrifugalpumpe, zusammen mit einer Heizeinheit und starten Bypass. </li><li> Schließen Sie die Isolierung der Leber durch Anziehen einer Rumel Stauschlauch an der Hohlvene infrahepatically über den Nierenvenen und Einspannen der Hohlvene suprahepatically mit einem Klintmalm Gefäßklemme. <ol><li> Ziehen Sie eine Rumel Stauschlauch an den gemeinsamen Gallengang, der Pfortader und der rechten Neben Vene und schließlich klemmen die Leberarterie mit einer Gefäßklemme. </li></ol></li></ol> 3. Die Perfusion <ol><li> Starten Sie die Durchblutung und den Blutspiegel im venösen Reservoir manuell überwachen. Die Durchflussmenge, bis ein konstantes Niveau erreicht ist, in der Regel eine Durchflussrate zwischen 500-1,200 ml / min. </li><li> Zeigen Thermistorsonden in die rechten und linken Leberlappen und einem direkt inder arterielle Katheter. </li><li> Erhitzen Sie das Perfusat, die auf eine Leberzieltemperatur von 40 o C. Halten Sie diese Temperatur während des Durchblutung. </li><li> Legen Sie eine Szintillationssonde über die venovenöse Bypasspumpe Kopf. </li><li> Kalibrieren des Leckageüberwachungssystems, durch Einspritzen einer kleinen Dosis von 10 MBq 99m Tc-Albumin (1 ml) in den systemischen Kreislauf und Registrieren der Grundlinienaktivität. <ol><li> Einprägen des zehnfach höheren Dosis, 100 MBq 99m Tc-Albumin (1 ml), in die Leberperfusion Einheit. </li><li> Nehmen ein Austreten aus dem Perfusionssystem in die systemische Zirkulation als Zunahme der Aktivität durch das Szintillations-Sonde gemessen. </li><li> Nehmen jegliche Leckage aus der systemischen Zirkulation zum Perfusionssystem als eine Volumenzunahme im venösen Reservoir. </li></ol></li><li> Beim stationären Zustand des Durchflusses eingerichtet ist, keine Leckage vorhanden ist und die Lebertemperatur 40 ° C erreicht hat, einzuspritzen Melphalan in einer Dosis von1 mg / kg Körpergewicht in das Perfusionssystem in zwei Dosen verabreicht 30 Minuten auseinander dividiert. </li></ol> 4. Beenden Sie die Perfusion und Katheter entfernen <ol><li> Nach 60 min der Perfusion, leeren Sie den Perfusat aus dem System und bewässern die Leber mit 1000 ml Kristalloide. </li><li> Fügen Sie eine Einheit von gepackten Erythrozyten an die Perfusion Einheit, ziehen lassen in die Leber und dann stoppen Sie die Durchblutung. </li><li> Lassen Sie die Klintmalm Klemme und die Rumel Aderpresse aus der Hohlvene. </li><li> Lassen Sie die Rumel Aderpresse aus dem Ligamentum hepatoduodenale. </li><li> Nehmen Sie die Unterbindung von der gastroduodenalen Arterie. Entfernen Sie den Katheter und Ligation des Gefäßes. </li><li> Lösen Sie die Klemme aus der Leberarterie. </li><li> Entfernen des Katheters aus gonadalen Vene und Ligieren des Gefäßes. </li><li> Stoppen Sie den venovenöse Bypasspumpe und ziehen Sie die Rohre. </li><li> Schließen Sie den Bauch mit einer ununterbrochenen Naht Faszien und dann eine intrakutane Hautnaht. </li><li> Entfernen Sie die catheters in den Oberschenkel und externen Jugularvenen und gelten lokalen Druck 5 Min. </li></ol> 5. Postoperative Behandlung <ol><li> Extubieren den Patienten und Transfer zum postoperativen Intensivstation für 24 Stunden. Halten Überwachung der zentrale Venendruck und arterielle Blutdruck. </li><li> Bringen Sie den Patienten an einen chirurgischen Abteilung für ein typisches von 3-7 Tagen nach der post-operative Betreuung. </li><li> Weiter mit Heparin mit niedrigem Molekular (LMWH) in einer Dosis von 5000 IE / Tag für insgesamt 4 Wochen. </li></ol>

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	<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.

Patienten mit metastasiertem Aderhautmelanom haben nur wenige Behandlungsmöglichkeiten. Chemotherapie 6, Immuntherapie 23 und zielgerichtete Therapie 24 nicht in der Lage, um eine verbesserte OS zeigen. Bei Patienten mit Lebermetastasen wenigen ist Leberresektion eine Option 12. Andere Behandlungen umfassen Leberintraarterieller Chemotherapie (HIA) 9, transarteriellen Chemoembolisation (TACE) 25 und selektive interne Radiotherapie (SIRT) 26. <...

Aderhautmelanom ist der häufigste primäre intraokulare Malignität in Erwachsene, die Inzidenz am höchsten in der kaukasischen Bevölkerung ein. Lokale Behandlung besteht in der Enukleation oder lokalen Plaque Strahlentherapie 2,3. Unabhängig von lokalen Behandlung wird Metastasen entwickeln schließlich in etwa 35-50% der Patienten 4,5. Die Leber ist die häufigste Stelle der Metastasen und etwa 50% der mit isolierten Lebermetastasen Patienten. Das mediane Gesamtüberleben (OS) für Patienten mit Lebermetastasen ist 6-12 Monate 4. Mehrere verschiedene systemische und regionale Behandlungsstrategien untersucht worden, aber die Überlebensraten haben sich nicht verbessert 2. Die Behandlung mit systemischen Chemotherapie, die beide Einzelsubstanz (Dacarbazin oder Temozolomid 6) oder Kombinationsbehandlungen (BOLD-Schema 7, Gemcitabin mit Treosulfan 8), zeigt Ansprechraten von weniger als10%. Leberintraarteriellen Chemotherapie (HIA) hat einen etwas höheren Gesamtreaktionsrate im Vergleich zur systemischen Chemotherapie, aber keine Erhöhung der Gesamtüberlebens 9 gezeigt. Transarteriellen Chemoembolisation (TACE) mit Cisplatin und Carboplatin zeigten eine partielle Remission in 57% der Patienten mit einer medianen Überlebens von 9 Monaten 10. Selektive interne Radiotherapie (SIRT) wurde eine partielle Remission in 62% der Patienten mit einer medianen Überlebens von 7 Monaten 11 gezeigt. In einer Studie zur Leberresektion, darunter 255 Patienten wurde ein Gesamtüberleben von 14 Monaten ausgewiesen; in 76 der Patienten, bei denen eine mikroskopisch vollständige Resektion möglich war, war die Überlebensrate 27 Monate 12. Isolierte Leberperfusion (IHP) ist eine regionale Behandlungsoption, die ursprünglich von Ryan und Ausman entwickelt. Die Logik hinter der Technik ist es, operativ zu isolieren und Perfusion der Leber mit einer hohen Dosis eines chemotherapeutischen Mittels unter Vermeidung systemischxicity 13. Darüber hinaus vermittelt die gleichzeitige Hyperthermie eine erhöhte Aufnahme des chemotherapeutischen Mittels durch Änderungen in der Tumordurchblutung und zellulärer Membranpermeabilität 14. Vollständige Gefäß Isolierung mit einem radioaktiven Tracer-Technik unter Verwendung von Tc-99 markierten menschlichen Serumalbumin 15 bestätigt. In einer retrospektiven Studie der Phase II hat eine mögliche Überlebensvorteil von 14 Monaten (26 vs. 12 Monate) bei Patienten, die IHP erlebte im Vergleich zu den längsten Überlebenden mit Aderhautmelanom Lebermetastasen in Schweden 16 gezeigt. Hier stellen wir die Technik des IHP mit einem kurzen Überblick über die aktuellen Ergebnisse und Zukunftsperspektiven.

<table><tbody><tr><td>Wire re-inforced catethers</td><td>Medtronic Inc, Minnesota, USA</td><td></td><td>12-18Fr</td></tr><tr><td>St&amp;ouml;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

Isolierte Leberperfusion (IHP) ist ein Verfahren, bei dem die Leber wird chirurgisch isoliert und mit einer hohen Konzentration des Chemotherapeutikums Melphalan perfundiert. Kurz gesagt, beginnt das Verfahren mit dem Aufbau einer perkutanen veno-venöse Bypass aus der Oberschenkelvene an die externe Halsschlagader. Über eine Laparotomie werden Katheter dann in die richtige Leberarterie und der Hohlvene eingesetzt. Die Pfortader und Hohlvene, sowohl supra- und infrahepatically, werden dann eingespannt. Die arterielle und venöse Katheter an eine Herz-Lungen-Maschine verbunden ist, und die Leber mit Melphalan (1 mg / kg Körpergewicht) für 60 min perfundiert. Auf diese Weise ist es möglich, lokal Perfusion der Leber mit einer hohen Dosis eines chemotherapeutischen Mittels, ohne Leckage in die systemische Zirkulation. In früheren Studien mit Patienten mit isolierter Lebermetastasen von Aderhautmelanom, eine allgemeine Ansprechrate von 33 bis 100% und eine mediane Überlebenszeit von 9 bis 13 Monate,berichtet worden. Ziel dieses Protokolls ist es, eine klare Beschreibung, wie man das Verfahren durchführen zu geben und IHP als Behandlungsoption für Lebermetastasen von Aderhautmelanom diskutieren.

Ein erster Bericht enthalten 22 Patienten mit Melphalan mit oder ohne TNF-alpha behandelt. Die Ergebnisse zeigten eine Gesamtansprechrate von 62%, davon zwei Patienten mit CR (10%) und eine mediane Überlebenszeit von 11 Monaten 20. Eine weitere Studie berichtet über die Ergebnisse der 29 aufeinander folgenden Patienten behandelt mit Melphalan allein. Das Ergebnis war sehr ähnlich mit 62% Gesamtansprechrate einschließlich 10% komplette Remission und eine mediane Überlebenszeit von 12 Monaten 21.</sup...

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.

Isolierte Leberperfusion als Behandlung für Lebermetastasen von Aderhautmelanom

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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Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

13.3K Aufrufe.  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.

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

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

Murine Modell der metastasierenden Lebertumoren in der Einstellung der Ischämie Reperfusionsverletzung

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Forschung

Krebsforschung

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more representative models to study human cancers in immunodeficient mice than traditional established human cell lines in vitro. Recently, orthotopically implanted patient-derived tumor xenograft (PDX) models in mice have been developed to better replicate features of patient tumors. A liver orthotopic xenograft mouse model is expected to be a useful cancer research platform, providing insights into tumor biology and drug therapy. However, liver orthotopic tumor implantation is generally complicated. Here we describe our protocols for the orthotopic implantation of patient-derived liver-metastatic uveal melanoma tumors. We cultured human liver metastatic uveal melanoma cell lines into immunodeficient mice. The protocols can result in consistently high technical success rates using either a surgical orthotopic implantation technique with chunks of patient-derived uveal melanoma tumor or a needle injection technique with cultured human cell line. We also describe protocols for CT scanning to detect interior liver tumors and for re-implantation techniques using cryopreserved tumors to achieve re-engraftment. Together, these protocols provide a better platform for liver orthotopic tumor mouse models of liver metastatic uveal melanoma in translational research.

Orthotopic human liver metastatic uveal melanoma xenograft mouse models were created using surgical orthotopic implantation techniques with patient-derived tumor chunk and needle injection techniques with cultured human uveal melanoma cell lines.

Generierung einer Leber orthotopischen Human Enuveal Melanom Xenograft Plattform bei immundefizienten Mäusen

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more ...

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

Portalveneninjektion von Darmkrebs-Organoiden zur Untersuchung der Lebermetastasierung Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for liver transplantation die without receiving an organ transplant or are delisted for disease progression. One strategy to increase the donor pool is the utilization of marginal grafts, such as fatty livers, grafts from older donors, or donation after cardiac death (DCD). The current preservation technique of cold static storage is only poorly tolerated by marginal livers resulting in significant organ damage. In addition, cold static organ storage does not allow graft assessment or repair prior to transplantation.
These shortcomings of cold static preservation have triggered an interest in warm perfused organ preservation to reduce cold ischemic injury, assess liver grafts during preservation, and explore the opportunity to repair marginal livers prior to transplantation. The optimal pressure and flow conditions, perfusion temperature, composition of the perfusion solution and the need for an oxygen carrier has been controversial in the past.
In spite of promising results in several animal studies, the complexity and the costs have prevented a broader clinical application so far. Recently, with enhanced technology and a better understanding of liver physiology during ex vivo perfusion the outcome of warm liver perfusion has improved and consistently good results can be achieved.
This paper will provide information about liver retrieval, storage techniques, and isolated liver perfusion in pigs. We will illustrate a) the requirements to ensure sufficient oxygen supply to the organ, b) technical considerations about the perfusion machine and the perfusion solution, and c) biochemical aspects of isolated organs.

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

Marginal grafts, such as fatty livers, grafts from older donors, or livers retrieved after cardiac death (DCD) tolerate conventional, cold static storage only poorly. We developed a novel model of subnormothermic ex vivo liver perfusion for preservation, assessment, and repair of marginal liver grafts prior to transplantation.

Technik der Subnormothermic<em&gt; Ex-vivo-</em&gt; Leberperfusion für die Lagerung, Bewertung, Reparatur von Marginal Lebertransplantate

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for ...

Medizin

Implantation and Evaluation of Melanoma in the Murine Choroid via Optical Coherence Tomography

Establishing experimental choroidal melanoma models is challenging in terms of the ability to induce tumors at the correct localization. In addition, difficulties in observing posterior choroidal melanoma in vivo limit tumor location and growth evaluation in real-time. The approach described here optimizes techniques for establishing choroidal melanoma in mice via a multi-step sub-choroidal B16LS9 cell injection procedure. To enable precision in injecting into the small dimensions of the mouse uvea, the complete procedure is performed under a microscope. First, a conjunctival peritomy is formed in the dorsal-temporal area of the eye. Then, a tract into the sub-choroidal space is created by inserting a needle through the exposed sclera. This is followed by the insertion of a blunt needle into the tract and the injection of melanoma cells into the choroid. Immediately after injection, noninvasive optical coherence tomography (OCT) imaging is utilized to determine tumor location and progress. Retinal detachment is evaluated as a predictor of tumor site and size. The presented method enables the reproducible induction of choroid-localized melanoma in mice and the live imaging of tumor growth evaluation. As such, it provides a valuable tool for studying intraocular tumors.

Implantation and Evaluation of Melanoma in the Murine Choroid via Optical Coherence Tomography

The present protocol describes the implantation and evaluation of melanoma in the murine choroid utilizing optical coherence tomography.

Implantation und Beurteilung des Melanoms in der Aderhaut der Maus mittels optischer Kohärenztomographie

Establishing experimental choroidal melanoma models is challenging in terms of the ability to induce tumors at the correct localization. In addition, ...

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.

Die perkutane Leberperfusion (PHP) mit Melphalan als Behandlung für inoperablen Metastasen auf die Leber beschränkt

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

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

Visualisierung von Gefäß- und Parenchymregeneration nach 70% Teilhepatektomie in normalen Mäusen

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

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.

Ein kleines Tier Modell der Ex Vivo Normothermic Leber Perfusion

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

Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

Partial hepatectomy has been widely used to investigate liver regeneration in mice, but the isolation of high yields of viable hepatocytes for downstream single-cell applications is challenging. A marked accumulation of lipids within regenerating hepatocytes is observed during the first 2 days of normal liver regeneration in mice. This so-called transient regeneration-associated steatosis (TRAS) is temporary but partially overlaps the major proliferative phase. Density-gradient purification is the backbone of most existing protocols for the isolation of primary hepatocytes. As gradient purification relies on the density and size of cells, it separates non-steatotic from steatotic hepatocyte populations. Therefore, fatty hepatocytes often are lost, yielding non-representative hepatocyte fractions. 
The presented protocol describes an easy and reliable method for the in vivo isolation of regenerating hepatocytes regardless of their lipid content. Hepatocytes from male C57BL/6 mice are isolated 24-48 h after hepatectomy by a classic two-step collagenase perfusion approach. A standard peristaltic pump drives the warmed solutions via the catheterized inferior vena cava into the remnant, using a retrograde perfusion technique with outflow through the portal vein. Hepatocytes are dissociated by collagenase for their release from the Glisson's capsule. After washing and careful centrifugation, the hepatocytes can be used for any downstream analyses. In conclusion, this paper describes a straightforward and reproducible technique for the isolation of a representative population of regenerating hepatocytes after partial hepatectomy in mice. The method may also aid the study of fatty liver disease.

Lipid-laden hepatocytes are inherent to liver regeneration but are usually lost upon density-gradient centrifugation. Here, we present an optimized cell isolation protocol that retains steatotic hepatocytes, yielding representative populations of regenerating hepatocytes after partial hepatectomy in mice.

Isolierung regenerierender Hepatozyten nach partieller Hepatektomie bei Mäusen

Partial hepatectomy has been widely used to investigate liver regeneration in mice, but the isolation of high yields of viable hepatocytes for downstream ...

Development and Validation of a Porcine Ex Situ Isolated Liver Perfusion Model 

Porcine Normothermic Isolated Liver Perfusion

Porcine models of liver ex situ normothermic machine perfusion (NMP) are increasingly being used in transplant research. Contrary to rodents, porcine livers are anatomically and physiologically close to humans, with similar organ size and bile composition. NMP preserves the liver graft at near-to-physiological conditions by recirculating a warm, oxygenated, and nutrient-enriched red blood cell-based perfusate through the liver vasculature. NMP can be used to study ischemia-reperfusion injury, preserve a liver ex situ before transplantation, assess the liver's function prior to implantation, and provide a platform for organ repair and regeneration. Alternatively, NMP with a whole blood-based perfusate can be used to mimic transplantation. Nevertheless, this model is labor-intensive, technically challenging, and carries a high financial cost.
In this porcine NMP model, we use warm ischemic damaged livers (corresponding to donation after circulatory death). First, general anesthesia with mechanical ventilation is initiated, followed by the induction of warm ischemia by clamping the thoracic aorta for 60 min. Cannulas inserted in the abdominal aorta and portal vein allow flush-out of the liver with cold preservation solution. The flushed-out blood is washed with a cell saver to obtain concentrated red blood cells. Following hepatectomy, cannulas are inserted in the portal vein, hepatic artery, and infra-hepatic vena cava and connected to a closed perfusion circuit primed with a plasma expander and red blood cells. A hollow fiber oxygenator is included in the circuit and coupled to a heat exchanger to maintain a pO2 of 70-100 mmHg at 38 &#176;C. NMP is achieved by a continuous flow directly through the artery and via a venous reservoir through the portal vein. Flows, pressures, and blood gas values are continuously monitored. To evaluate the liver injury, perfusate and tissue are sampled at predefined time points; bile is collected via a cannula in the common bile duct.

The porcine model of liver normothermic machine perfusion (NMP), described here, can be successfully used to study NMP as a preservation strategy, a tool for viability assessment, and a platform for organ repair. It holds a high translational value, however it is technically challenging and labor-intensive.

Porcine models of liver ex situ normothermic machine perfusion (NMP) are increasingly being used in transplant research. Contrary to rodents, porcine ...