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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