Name: re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy
Uploaded: 2018-04-08T13:00:00
Description: Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

This research was supported by National Natural Science Foundation of China (Grant No. 81501382) and Zhejiang Provincial Natural Science Foundation of China (Grant No. LQ13H100003 and LQ16H100002).

All of the procedures followed the guidelines of rodent surgery approved by the Wenzhou Medical University Animal Policy and Welfare Committee 15.
1. Animals
<ol>
	<li>Use male Lewis rats weighing 250-350 g as donors and recipients.</li>
</ol>
2. Operative Environment
<ol>
	<li>Perform all microsurgical procedures under a surgical microscope with a magnification ranging from 7X to 20X. Sterilize all surgical instruments before use in operatio...

<ol>
	<li>Yan, J. Q., Becker, T., Peng, C. H., Li, H. W., Klempnauer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Split+liver+transplantation:+a+reliable+approach+to+expand+donor+pool.">Split liver transplantation: a reliable approach to expand donor pool.</a> Hepatobiliary Pancreat Dis Int. 4, 339-344 (2005).</li><li>Xia, R., Emond, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+partial+liver+transplantation+in+the+rat:+a+model+of+70%+hepatectomy+and+reduced+size+liver+transplantation.">Orthotopic partial liver transplantation in the rat: a model of 70% hepatectomy and reduced size liver transplantation.</a> Transplantation. 56, 1041-1043 (1993).</li><li>Omura, T., Ascher, N. L., Emond, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fifty-percent+partial+liver+transplantation+in+the+rat.">Fifty-percent partial liver transplantation in the rat.</a> Transplantation. 62, 292-293 (1996).</li><li>Foss, 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=Function+of+reduced-size+liver+transplant+in+the+rat.">Function of reduced-size liver transplant in the rat.</a> Res Exp Med (Berl). 197, 91-99 (1997).</li><li>Nagai, K., Yagi, S., Uemoto, S., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+procedures+for+a+rat+model+of+partial+orthotopic+liver+transplantation+with+hepatic+arterial+reconstruction.">Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction.</a> J Vis Exp. , e4376 (2013).</li><li>Sun, J., Qin, G., Wu, L., Wang, C., Sheil, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigenic+load+and+peripheral+chimeric+levels+in+entire+and+partial+liver+allograft+recipients.">Antigenic load and peripheral chimeric levels in entire and partial liver allograft recipients.</a> Microsurgery. 21, 183-187 (2001).</li><li>Ninomiya, 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=Deceleration+of+regenerative+response+improves+the+outcome+of+rat+with+massive+hepatectomy.">Deceleration of regenerative response improves the outcome of rat with massive hepatectomy.</a> Am J Transplant. 10, 1580-1587 (2010).</li><li>Yamanaka, K., 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+olprinone+on+liver+microstructure+in+rat+partial+liver+transplantation.">Effect of olprinone on liver microstructure in rat partial liver transplantation.</a> J Surg Res. 183, 391-396 (2013).</li><li>Wang, W., 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=Mesenchymal+stem+cells+promote+liver+regeneration+and+prolong+survival+in+small-for-size+liver+grafts:+involvement+of+C-Jun+N-terminal+kinase,+cyclin+D1,+and+NF-kappaB.">Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts: involvement of C-Jun N-terminal kinase, cyclin D1, and NF-kappaB.</a> PLoS One. 9, e112532 (2014).</li><li>Madrahimov, N., Dirsch, O., Broelsch, C., Dahmen, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marginal+hepatectomy+in+the+rat:+from+anatomy+to+surgery.">Marginal hepatectomy in the rat: from anatomy to surgery.</a> Ann. Surg. 244, 89-98 (2006).</li><li>Martins, P. N., Theruvath, T. P., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+partial+hepatectomies.">Rodent models of partial hepatectomies.</a> Liver Int. 28, 3-11 (2008).</li><li>Kamada, N., Sumimoto, R., Kaneda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+value+of+hepatic+artery+reconstruction+as+a+technique+in+rat+liver+transplantation.">The value of hepatic artery reconstruction as a technique in rat liver transplantation.</a> Surgery. 111, 195-200 (1992).</li><li>Steger, U., Sawitzki, B., Gassel, A. M., Gassel, H. J., Wood, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+hepatic+rearterialization+on+reperfusion+injury+and+outcome+after+mouse+liver+transplantation.">Impact of hepatic rearterialization on reperfusion injury and outcome after mouse liver transplantation.</a> Transplantation. 76, 327-332 (2003).</li><li>Huang, 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=A+novel+end-to-side+anastomosis+technique+for+hepatic+rearterialization+in+rat+orthotopic+liver+transplantation+to+accommodate+size+mismatches+between+vessels.">A novel end-to-side anastomosis technique for hepatic rearterialization in rat orthotopic liver transplantation to accommodate size mismatches between vessels.</a> Eur Surg Res. 47, 53-62 (2011).</li><li>Bernal, J., 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=Guidelines+for+rodent+survival+surgery.">Guidelines for rodent survival surgery.</a> J.Invest Surg. 22, 445-451 (2009).</li><li>Johnson, A. J., Stulting, R. D., Macsai, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ophthalmic Microsurgical Suturing Techniques. , 21-28 (2007).</li><li>Delriviere, L., 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=Detailed+modified+technique+for+safer+harvesting+and+preparation+of+liver+graft+in+the+rat.">Detailed modified technique for safer harvesting and preparation of liver graft in the rat.</a> Microsurgery. 17, 690-696 (1996).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp. , (2012).</li><li>Delriviere, L., 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=Technical+details+for+safer+venous+and+biliary+anastomoses+for+liver+transplantation+in+the+rat.">Technical details for safer venous and biliary anastomoses for liver transplantation in the rat.</a> Microsurgery. 18, 12-18 (1998).</li><li>Jin, 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=Preliminary+experience+of+a+PDCA-cycle+and+quality+management+based+training+curriculum+for+rat+liver+transplantation.">Preliminary experience of a PDCA-cycle and quality management based training curriculum for rat liver transplantation.</a> J Surg Res. 176, 409-422 (2012).</li><li>Selzner, N., Selzner, M., Tian, Y., Kadry, Z., Clavien, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cold+ischemia+decreases+liver+regeneration+after+partial+liver+transplantation+in+the+rat:+A+TNF-alpha/IL-6-dependent+mechanism.">Cold ischemia decreases liver regeneration after partial liver transplantation in the rat: A TNF-alpha/IL-6-dependent mechanism.</a> Hepatology. 36, 812-818 (2002).</li><li>Tanaka, H., Hashizume, K., Enosawa, S., Suzuki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+a+20%+partial+liver+graft+in+rats:+a+technical+innovation.">Successful transplantation of a 20% partial liver graft in rats: a technical innovation.</a> J Surg Res. 110, 409-412 (2003).</li><li>Bockhorn, 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=Erythropoietin+treatment+improves+liver+regeneration+and+survival+in+rat+models+of+extended+liver+resection+and+living+donor+liver+transplantation.">Erythropoietin treatment improves liver regeneration and survival in rat models of extended liver resection and living donor liver transplantation.</a> Transplantation. 86, 1578-1585 (2008).</li><li>Martins, P. N., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+anatomy+of+the+liver,+hepatic+vasculature+and+bile+ducts+in+the+rat.">Surgical anatomy of the liver, hepatic vasculature and bile ducts in the rat.</a> Liver Int. 27, 384-392 (2007).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Kashfi, 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=A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplant.Proc. 37, 185-188 (2005).</li><li>Wain, R. A. J., Hammond, D., McPhillips, M., Whitty, J. P. M., Ahmed, W., Ahmed, W., Jackson, M. 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Rat PLTx is a sophisticated microsurgical procedure with a training program that is high in cost and long in duration20. The complexity of the rat PLTx protocol has prevented researchers from using this animal model. Compared with full-size rat LTx, rat PLTx presents the microsurgeon not only with the challenge of a transplantation procedure but also with the challenge of liver resection in a small animal. Therefore, a visualized microsurgical protocol describing the whole procedure in detail is e...

Currently, there is a large discrepancy between the number of donated liver organs and the number of patients waiting for a donated liver. The shortage of liver organs is a global problem. To expand the donor pool, split liver transplantation (LTx) and living donor LTx were developed to use a partial liver as a graft1.
To further investigate the mechanism behind partial liver transplantation (PLTx), relevant animal models have been established2,3,4,5. In rat PLTx, the liver lobes are resected in vivo and ex vivo to mimic the conditions of the living donor LTx and split LTx, respectively, in human. A paper published in the Journal of Visualized Experiments presented a detailed protocol involving a 50% rat PLTx using an ex vivo hepatectomy5. However, a rat PLTx with an in vivo hepatectomy has not yet been reported in the visualized literature.
In addition to the difference between in vivo and ex vivo hepatectomies, the technique of performing a hepatectomy itself also plays an important role in determining the outcome of PLTx. Currently, in many surgical studies using the rat PTLx model, the liver lobes were resected after placing a simple ligation at the pedicle of the liver lobe2,3,6,7,8,9. However, placing a simple ligation before resection is not suitable for all liver lobes, as different liver lobes have different shapes and sizes. A simple ligation at the base of the median lobe carries a high risk of causing a constriction of the vena cava, which might eventually affect the outflow of the partial liver graft10,11. Therefore, an update hepatectomy technique based on knowledge of the rat hepatic anatomy is required in the field of rat PTLx.
In the protocol described in this study, an updated vessel-oriented 70% hepatectomy was incorporated into the procedure of the rat PLTx. The portal veins and hepatic arteries of the left lateral lobe (LLL) and median lobe (ML) were dissected and divided individually before removal of the liver parenchyma. Then, the hepatic veins of the LLL and ML were ligated by piercing sutures. By using individual ligations and multiple piercing sutures rather than placing a simple ligation, the remnant stump of the ML was able to spread over the vena cava. Hence, the constriction of the infrahepatic vena cava caused by a surgical ligation was avoided. Additionally, occlusion of the blood supply of the LLL and ML by individual ligations before removal of the liver parenchyma decreased the rate of bleeding in the remnant liver stump, thereby minimizing the influence of blood loss on the experiments11.
For microsurgeons, it is a significant challenge to reconstruct the proper hepatic artery (PHA) of a liver graft because of the extremely small diameter of this vessel. Although the question of whether re-arterialization in LTx is truly necessary is still under debate12,13,14, numerous microsurgical techniques for reconstructing the hepatic artery have been proposed14. Here, we introduce a novel technique for re-arterialization of the liver graft, which anastomoses the common hepatic artery (CHA) to the enlarged PHA in an end-to-side manner. By using this end-to-side technique, two hepatic arteries are connected with an anastomosis of a larger diameter. The larger the diameter of the anastomosis is, the easier hand suturing is to perform and the greater the improvement in the patency of the anastomosis becomes.

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		<tr>
			<td>Surgical microscope</td>
			<td>M&#246;ller-Wedel, Germany</td>
			<td>654359</td>
			<td></td>
		</tr>
		<tr>
			<td>Light source</td>
			<td>OSRAM (China) Lighting Co., Ltd</td>
			<td>64627</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane Vaporizer</td>
			<td>MIDMARK Corporation</td>
			<td>VIP3000</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Baxter Healthcare Corporation</td>
			<td>40032609</td>
			<td>For Inhalation anesthesia</td>
		</tr>
		<tr>
			<td>Normal Saline</td>
			<td>Zhejiang Tianrui Pharmaceutical Co., Ltd.</td>
			<td>716092103</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone Iodine Solution</td>
			<td>HANGZHOU MINSHENG PHARMACEUTICAL</td>
			<td>H33021567</td>
			<td>For disinfection</td>
		</tr>
		<tr>
			<td>Hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J31020</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>JC2116</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle-holder</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J32030</td>
			<td></td>
		</tr>
		<tr>
			<td>Dressing Forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>J42020</td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>W40340</td>
			<td>For liver resection; For retracting the stay sutures.&#160;</td>
		</tr>
		<tr>
			<td>Micro-scissors</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA1040</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro needle-holder</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA2060</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved micro typing forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3061</td>
			<td></td>
		</tr>
		<tr>
			<td>straight micro typing forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3060</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro hemostatic forceps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>WA3020</td>
			<td></td>
		</tr>
		<tr>
			<td>Bulldog clamp</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>XEC350</td>
			<td></td>
		</tr>
		<tr>
			<td>Vessel clamps</td>
			<td>Shanghai Medical Instruments(Group) Ltd</td>
			<td>W40160</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Serrefine-curved</td>
			<td>&#160;Fine Science Tools, Inc</td>
			<td>18055-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Dacryosyrinx</td>
			<td>Shi Zhuo medical instruments Co., Ltd.</td>
			<td>5# curve</td>
			<td>Use as the curved needle for forcing out the air bubbles in the vascular lumen</td>
		</tr>
		<tr>
			<td>Sterilized gauze swabs</td>
			<td>Fuqing Health &#38; Integral Medical</td>
			<td>20230R</td>
			<td></td>
		</tr>
		<tr>
			<td>5mL syringe</td>
			<td>Zhejiang Yusheng Medical Instrument Co.,Ltd</td>
			<td>811932416</td>
			<td></td>
		</tr>
		<tr>
			<td>18G needles</td>
			<td>Shanghai Kindly Enterprise Development Group Co., Ltd</td>
			<td>01-014</td>
			<td>For fixing the upper abdomen wall</td>
		</tr>
		<tr>
			<td>Intima II 22G intraveous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>383207y</td>
			<td>For making biliary stent; For infusing perfusate</td>
		</tr>
		<tr>
			<td>7-0 polypropylene sutures</td>
			<td>Ningbo Medical Needle Co., Ltd</td>
			<td>151026</td>
			<td>For SHVC anastomsis</td>
		</tr>
		<tr>
			<td>6-0 silk suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products Co., Ltd</td>
			<td>2650182</td>
			<td>For ligation</td>
		</tr>
		<tr>
			<td>Culture dish (100mm)</td>
			<td>Corning Incorporated</td>
			<td>430165</td>
			<td>used in back table for storage ice. The back table dish was placed on it.&#160;</td>
		</tr>
		<tr>
			<td>Culture dish (30 mm)</td>
			<td>Corning Incorporated</td>
			<td>3296</td>
			<td>As a vessel for organ preservation</td>
		</tr>
		<tr>
			<td>Angiocath 12G intravenous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>382277</td>
			<td>For making a IHVC cuff</td>
		</tr>
		<tr>
			<td>Angiocath 14G intravenous catheter</td>
			<td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td>
			<td>382269</td>
			<td>For making a PV cuff</td>
		</tr>
		<tr>
			<td>Buprenorphine (hydrochloride)</td>
			<td>Tianjin Institute of Pharmaceutical Research Pharmaceutical Co., Ltd</td>
			<td>H12020275</td>
			<td>For analgesic</td>
		</tr>
		<tr>
			<td>Cefuroxime sodium for injection</td>
			<td>Esseti FarmaceuticiS.r.l</td>
			<td>H20160013</td>
			<td>As an antibiotics</td>
		</tr>
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re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

In total, 31 cases of syngeneic arterialized rat PLTx were completed using this protocol. All of the recipients survived until the end of the observation time. The body weight of the recipients began to recover after postoperative day (POD) 4. The slope of the body weight was close to that of a normal Lewis rat after POD 6 (Figure 8), which indirectly implied the recovery of the recipient. Histologically, a slight proliferation of the bile duct was observed i...

Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

A protocol involving re-arterialized rat partial liver transplantation is presented in this article. Specifically, 70%of liver was resected in vivo by using the updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner.The cuff technique was modified to shorten the anastomosis time of the inferior patic vena cava. Isoflurane mixed with zero point five liter per minute oxygen was used to induce and maintain inhalation anesthesia. Cut a small transverse incision on the upper abdomen.Then, extend the transverse incision close to the costal margin on the upper abdomen. Reverse and affix upper abdomen wall to the cranial side. Next, reverse the xiphoid process to cranial side, too.Cut off the falciform ligament, coronary ligament, and a triangular ligament, in sequence. Free the left diaphragmatic vein from the diaphragm using microforceps. Then, double ligate it using six-zero silk sutures.Free the hepatoesophageal ligament and artery. Then, double-ligate it and divide it. Expose the first hepatic hilum of the liver.First, carefully dissect the posterior wall of the common trunk of the portal vein. Then, double ligate it and divide it with the corresponding bile duct and the hepatic artery together. Second, gently dissect, double-ligate and divide the portal vein of the right medium lobe.Third, double ligate and divide the hepatic artery and the bile duct of the right medium lobe using six-zero silk sutures. Ligate the hepatic of the left lateral lobe using circumstantial three-zero silk suture. Remove the liver mass using micro scissors above the ligation.Clamp the left median hepatic vein by mosquito hemostatic forceps placed along the median fissure. Remove all of the liver parenchyma above the forceps using micro scissors. Place the piercing suture under the forceps surrounding the left median hepatic vein.Then, tie a square knot to ligate it. Besides, place another two piercing sutures to close the rest of the incision. After this, place a mosquito hemostatic forceps at a distance of point two-five centimeters to occlude the right median hepatic vein.Remove all liver parenchyma above the forceps. Place the first piercing suture penetrating the liver parenchyma under the forceps to surround the middle median hepatic vein. Then, tie a square knot to ligate it.Place a second piercing suture under the forceps to surround the middle median hepatic vein. Then, ligate it using the same method. Place another two piercing sutures under the mosquito hemostatic forceps to close the rest of the incision of the medium lobe by using these piercing sutures.The remnant stump of the medium lobe was able to spread over the vena cava. Cut open the retroperitoneum to expose the inferior hepatic vena cava. Dissect and ligate the right adrenal vein, which drains into the inferior hepatic vena cava from the rear.Free the right renal artery and the right renal vein. Perform a right nephrectomy after ligating the right renal vein and the right renal artery. Mobilize the common bile duct from the first hepatic hilum using micro forceps.Then, ligate the common bile duct at the crest point of the polaric vein and the common bile duct. Cut a transverse incision on the interior wall of the common bile duct. Insert a 22-gauge biliary stent immediately into lumen of the common bile duct.Secure the biliary stent with circumferential six-zero silk suture. Transect the common bile duct between the two six-zero silk sutures. Free the pyloric vein using micro forceps then double ligate and divide it.Next, double ligate and divide the splenic vein the same way. Carefully free the common hepatic artery and it's two bifurcations. Double ligate the gastroduodenal artery using six-zero silk sutures at the bifurcation of the common hepatic artery to gastroduodenal artery.Inject 50 IU of heparin in one milliliter of saline via the penile vein. Cross-clamp the inferior hepatic vena cava portal vein and the common hepatic artery using three curved micro Perform a small incision on the interior wall of the portal vein to insert a 22-gauge catheter. Profuse the partial liver graft with saline at four degrees celsius via the catheter under pressure of 20 centimeters water.Rapidly cut open the thoracic cavity and intrathoracic vena cava using surgical scissors to drain out the blood and profuse it Use micro scissors to transect the left diaphragmatic vein between the two ligations which were placed previously. Next, transect the suprahepatic vena cava close to the diaphragm to keep the vascular lanes as long as possible. Cut off the portal vein at the level of the splenic vein and transect the common hepatic artery close to the select trunk.Cut off the adrenal vessels on the distill side of the ligation. Then, divide the infrahepatic vena cava at the level of the left renal vein. Pull the portal vein through the lumen of the cuff made with a 12-gauge intravenous catheter.Temporarily fix puff handle and portal vein together. Avert the portal vessel wall to cover the outside surface of the cuff. Secure the vessel wall and the cuff in position with circumferential six-zero silk suture.Install the infrahepatic vena cava cuff using the same method. Penetrate the vessel wall of the suprahepatic vena cava from outside to inside using two seven-zero polypropylene sutures along the three o'clock and nine o'clock directions, respectively. Repeat the procedures in 6.1, 6.2, and 6.7 in the recipient, except for the right nephrectomy.Mobilize, double ligate, and divide the common bile duct at the first bifurcation in the hepatic hilum. Transect and prepare the hepatic artery using micro scissors, subsequent to double ligation using six-zero silk suture. Retract the recipient liver to the left side using two cotton sticks.Cut off the ligament in the rear of the liver from the suprahepatic vena cava to the right adrenal vein. Introduce a rubber band through the back space of the suprahepatic vena cava. Cross-clamp the infrahepatic vena cava above the renal vein using a vessel clamp and the portal vein above the pyloric vein.the suprahepatic vena cava and diaphragm and clamp the parts of the diaphragm together with the intrathoracic vena cava. Cut the vessel wall of the suprahepatic vena cava carefully along the upper edge of the liver parenchyma to keep the vascular wall as long as possible. Transect the portal vein at the first bifurcation in the hepatic hilum using micro scissors.Transect the inferior vena cava and parenchyma of the right inferior lobe rather than in the vessel wall to leave a section of circumferential liver parenchyma around the vena cava. Keep the cutting edge in the right inferior lobe at a distance of four millimeters to the inferior hepatic vena cava. Place the partial liver graft in the abdominal cavity, isotopically.Pierce the vessel wall of the suprahepatic vena cava from inside to outside along three o'clock and nine o'clock directions Using two prestat stay sutures. Pull the stay sutures to the right and left after tying into a knot with their own ends. Pierce the suprahepatic vena cava vessel wall from outside to inside, close to the knot in the three o'clock direction.Anastomose the posterior wall of the suprahepatic vena cava by running suture inside the vascular lumen from the three o'clock direction to the nine o'clock direction. At the corner of the nine o'clock direction, pierce the vessel wall from inside to outside, close to the knot, to introduce the needle out of the vascular lumen. Use this suture to anastomose interior wall using running suture outside the vascular lumen from the nine o'clock direction to the three direction.Finally, tie the suture with its own end three times. Place two seven-zero polypropylene stay sutures at the end of the portal vein of the recipient, following the description in seven point two. Pull two stay sutures in the opposite direction to extend the vessel wall.Inject approximately one milliliter of saline into the vascular lumen of the portal vein to force out air bubbles. Quickly insert the portal vein cuff into the vascular lumen of the portal vein in the recipient when it is opened using microforceps. Secure the cuff anastomosis using circumferential six-zero silk suture.Then, release the clamps on the suprahepatic vena cava and the portal vein, in sequence. Inject one milliliter of saline into the vascular lumen to force out air bubbles. Insert the cuff into cylindrical vascular lumen of the inferior hepatic vena cava.Quickly secure the cuff anastomosis using a six-zero silk suture. Release the two vessel clamps in the inferior hepatic vena cava to restart the profusion. Then, trim off the surrounding liver parenchyma carefully, above the circumferential silk suture using micro scissors.Penetrate the vessel wall of the common hepatic artery in the liver graft from outside to inside using two eleven-zero polypropylene sutures in the three o'clock and nine o'clock directions, respectively. Occlude the blood flow of the common hepatic artery and the gastroduodenal artery in the recipient using two curved micro Transect the proper hepatic artery of the recipient at its root to expose the vascular lumen. Enlarge the vascular lumen of the proper hepatic artery in the recipient by cutting part of the vessel wall, longitudinally, to accommodate the diameter of the common hepatic artery in the liver graft.Flush the vascular lumen using saline. Anastomose the common hepatic artery of the liver graft to enlarge the proper hepatic artery in an end-to-side manner using the running suture technique. Perform a transverse incision on the anterior wall of the bile duct in the recipient.Pull the biliary stent in the partial liver graft down to insert it into the lumen of the bile duct in the recipient. Divide the transverse incision. Secure the stent using six-zero circumferential silk suture.Tie the two six-zero circumferential silk sutures in the bile duct to each other to reduce the tension of the anastomosis. After surgery, treat all recipients with 0.1 milligram per kilogram Buprenorphine and Cefuroxime 16 milligram per kilogram, subcutaneously. In total, 31 cases have seen generic arterialized rat partial liver transplantation were completed using this protocol.All of the recipients survived until the end of the observation time. The body weight of the recipient began to recover after postoperative day four. The slope of the body weight was close to that of the Lewis rat after postoperative day six, which indirectly implied recovery of the recipient.Histologically, a slight proliferation of the bile duct was observed in the recipients and the liver lobular architecture of the recipient was intact. No necrosis or obvious sign of pseudo dilation was observed. The serum level of alanine aminotransferase was in the normal range at postoperative day 30.No cases of yellow urine caused by jaundice were observed in these recipients, which was indicated by checking the cage bedding daily, indicating the patency of the biliary stent. The normal serum level of bilirubin on postoperative day 30 further confirmed the success for reconstruction of the bile duct. The average duration of the anhepatic phase was 22.03 plus or minus 2.3 minutes.The cuff anastomosis of the portal vein took 4.68 plus or minus 0.77 minutes, while the cuff anastomosis of the inferior hepatic vena cava only took 2.05 plus or minus 0.71 mminutes. Comparatively, the cuff anastomosis of the portal vein, the cuff anastomosis of the inferior hepatic vena cava saved the staff of placing stay sutures, and thereby, further shortens the anastomosis time. Two cases of bleeding in the remnant stump of the medium lobe during the 70%hepatectomy were observed, among 31 cases of vessel-oriented hepatectomy.In the previous experiment, bleeding in the remnant stump of the medium lobe was observed in eight out of 18 cases of parenchyma, preserving vessel-oriented hepatectomy. This result indicated a lower rate of bleeding in the remnant liver stump of the medium lobe, after using vessel-oriented 70%hepatectomy. By using this protocol of rat partial liver transplantation, a low liver enzyme level and intact liver lobular architecture and a high survival rate were achieved after microsurgery.

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Research

JoVE Journal

Medicine

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

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.

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

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

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.

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

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological conditions. Partial hepatectomy also facilitates the increased engraftment and proliferation of transplanted cells by accelerating neovascularization and cell migration towards the liver. Here, we describe a simple protocol for performing 30% hepatectomy and transplantation of cells in the spleen of a non-obese diabetic/severe combined immunodeficient NOD.SCID (NOD.CB17-Prkdcscid/J) mouse. 
In this procedure, two small incisions are made. The first incision is to expose and resect the left lobe of the liver, and another small incision is made to expose the spleen for the intrasplenic transplantation of cells. This procedure does not require any specialized surgical skills, and it can be completed in 5-7 minutes with less stress and pain, faster recovery, and better survival. We have demonstrated the transplantation of hepatocytes isolated from a green fluorescent protein (GFP) expressing mouse (Transgenic C57BL/6-Tg (UBC-GFP) 30Scha/J), as well as hepatocyte like cells of human origin (NeoHep) in partially hepatectomized NOD.SCID mice.

We have described a protocol for performing partial hepatectomy (PHx) and cell transplantation via spleen in NOD.SCID (NOD.CB17-Prkdcscid/J) mice. In this protocol, an incision is made to expose and resect the left lobe of the liver followed by another incision for the intrasplenic transplantation of cells.

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

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.

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