Name: Author Spotlight: Advancing Liver Regeneration Research through ALPPS Mouse Model
Uploaded: 2024-01-19T14:00:00
Description: Read this Scientific Article Protocol about A Mouse Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Procedure Aided by Microscopy at JoVE.com

All experiments in this protocol were approved by the Veterinary Authorities of the Jiangxi Provincial People&#39;s Hospital (number 70/2022). Furthermore, all experimental steps were performed in strict compliance with the Institutional Animal Care and Use Committee.
1. Start of surgery
NOTE: Male C57BL/6 mice weighing 20-22 g were housed in a well-ventilated cage, maintained in a standard pathogen-free environment with a 12 h/12 h light/dark cycle. ...

Studying Liver Regeneration of ALLPS in Animal Models for Staged Hepatectomy

<ol>
	<li>Michalopoulos, G. K., Bhushan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+regeneration:+Biological+and+pathological+mechanisms+and+implications.">Liver regeneration: Biological and pathological mechanisms and implications.</a> Nat Rev Gastroenterol Hepatol. 18 (1), 40-55 (2021).</li><li>Chapelle, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+the+future+remnant+liver+function+is+a+better+tool+to+pr+edict+post-hepatectomy+liver+failure+than+platelet-based+liver+scores.">Estimation of the future remnant liver function is a better tool to pr edict post-hepatectomy liver failure than platelet-based liver scores.</a> Eur J Surg Oncol. 43 (12), 2277-2284 (2017).</li><li>Isfordink, C. 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=Portal+vein+ligation+versus+portal+vein+embolization+for+induction+of+hypertrophy+of+the+future+liver+remnant:+A+systematic+review+and+meta-analysis.">Portal vein ligation versus portal vein embolization for induction of hypertrophy of the future liver remnant: A systematic review and meta-analysis.</a> Surg Oncol. 26 (3), 257-267 (2017).</li><li>Linecker, 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=Risk+adjustment+in+alpps+is+associated+with+a+dramatic+decrease+in+ear+ly+mortality+and+morbidity.">Risk adjustment in alpps is associated with a dramatic decrease in ear ly mortality and morbidity.</a> Ann Surg. 266 (5), 779-786 (2017).</li><li>Darnis, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subtotal+hepatectomy+in+swine+for+studying+small-for-size+syndrome+and+portal+inflow+modulation:+Is+it+reliable.">Subtotal hepatectomy in swine for studying small-for-size syndrome and portal inflow modulation: Is it reliable.</a> HPB (Oxford). 17 (10), 881-888 (2015).</li><li>Casellas, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inbred+mouse+strains+and+genetic+stability:+A+review.">Inbred mouse strains and genetic stability: A review.</a> Animal. 5 (1), 1-7 (2011).</li><li>Zhang, 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=Evolution+of+associating+liver+partition+and+portal+vein+ligation+for+staged+hepatectomy+from+2012+to+2021:+A+bibliometric+analysis.+Review.">Evolution of associating liver partition and portal vein ligation for staged hepatectomy from 2012 to 2021: A bibliometric analysis. Review.</a> Int J Surg. 103, 106648 (2022).</li><li>Parola, M., Pinzani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+fibrosis:+Pathophysiology,+pathogenetic+targets+and+clinical+iss+ues.">Liver fibrosis: Pathophysiology, pathogenetic targets and clinical iss ues.</a> Mol Aspects Med. 65, 37-55 (2019).</li><li>Kikuchi, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+associating+liver+partition+and+portal+vein+occlusion+for+st+aged+hepatectomy+on+tumor+growth+in+a+mouse+model+of+liver+metastasis.">Impact of associating liver partition and portal vein occlusion for st aged hepatectomy on tumor growth in a mouse model of liver metastasis.</a> Eur J Surg Oncol. 44 (1), 130-138 (2018).</li><li>Schlegel, 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=Alpps:+From+human+to+mice+highlighting+accelerated+and+novel+mechanism+s+of+liver+regeneration.">Alpps: From human to mice highlighting accelerated and novel mechanism s of liver regeneration.</a> Ann Surg. 260 (5), 839-846 (2014).</li><li>Dili, A., Lebrun, V., Bertrand, C., Leclercq, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Associating+liver+partition+and+portal+vein+ligation+for+staged+hepate+ctomy:+Establishment+of+an+animal+model+with+insufficient+liver+remnan+t.">Associating liver partition and portal vein ligation for staged hepate ctomy: Establishment of an animal model with insufficient liver remnan t.</a> Lab Invest. 99 (5), 698-707 (2019).</li><li>Schadde, E., Hertl, M., Breitenstein, S., Beck-Schimmer, B., Schl&#228;pfer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+model+of+the+associating+liver+partition+and+portal+vein+ligation+for+staged+hepatectomy+(alpps)+procedure.">Rat model of the associating liver partition and portal vein ligation for staged hepatectomy (alpps) procedure.</a> J Vis Exp. (126), e55895 (2017).</li><li>Allaire, 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=New+frontiers+in+liver+resection+for+hepatocellular+carcinoma.">New frontiers in liver resection for hepatocellular carcinoma.</a> JHEP Rep. 2 (4), 100134 (2020).</li><li>Memi, F., Ntokou, A., Papangeli, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crispr/cas9+gene-editing:+Research+technologies,+clinical+applications+and+ethical+considerations.">Crispr/cas9 gene-editing: Research technologies, clinical applications and ethical considerations.</a> Semin Perinatol. 42 (8), 487-500 (2018).</li><li>Saviano, A., Henderson, N. C., Baumert, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+genomics+and+spatial+transcriptomics:+Discovery+of+novel+cell+states+and+cellular+interactions+in+liver+physiology+and+disease+biology.">Single-cell genomics and spatial transcriptomics: Discovery of novel cell states and cellular interactions in liver physiology and disease biology.</a> J Hepatol. 73 (5), 1219-1230 (2020).</li></ol>

This study presents a protocol for the initial phase of ALPPS surgery in mice, involving partial ligation of the portal vein and division of the median liver lobe. This method closely mimics the human ALPPS procedure, with the mouse liver&#39;s unique lobular structure and dual portal venous supply to the median lobe ensuring distinct blood flow post-surgery11. It is noteworthy that the ALPPS procedure comprises two distinct surgical stages. In this study, we exclusively present the initial stage ...

The liver harbors formidable regenerative potential, swiftly proliferating and restoring metabolic demands within a mere 3 months following resection for diverse hepatic ailments1. However, the imperative to ascertain the completeness of tumor margins necessitates the inevitability of expansive hepatic excision. Thus, ensuring an ample volume of the participatory hepatic milieu, known as the future liver remnant (FLR), assumes paramount importance2. ALPPS has been a breakthrough technique in hepatic surgery over the past few decades, particularly tailored for patients with inadequate residual hepatic volume subsequent to tumor resection, heralded as one of the most auspicious breakthroughs in the realm of hepatic oncological surgery3.
Remarkable progress has been made in developing ALPPS animal models. An ideal model typically requires independent blood inflow (portal vein and hepatic artery) and outflow (hepatic vein) in the preserved hepatic lobe and a clear separation between the preserved and the to-be-resected hepatic lobes to prevent collateral circulation4. Although ALPPS stimulates rapid hepatic regeneration in the remaining liver tissue, the specific mechanisms of this process are still unclear.
Currently, ALPPS models are categorized into three types: large animal models (e.g., swine and sheep), medium-sized models (e.g., rabbits and rodents), and small models (e.g., mice)5. The use of mice, with their quick breeding and ease of genetic modification, is particularly effective for in-depth studies of hepatic regeneration mechanisms6. Moreover, the liver structure of mice, especially their middle hepatic vein, closely resembles that of humans, making them highly suitable for ALPPS model development.
It is important to note that most hepatocellular carcinoma patients in clinical practice have underlying liver conditions, unlike the healthy liver models typically used in studies7. Thus, using mice preconditioned with hepatic fibrosis or viral infections can more accurately simulate the surgical responses and postoperative liver regeneration seen in patients with various liver diseases8. This approach could reveal new therapeutic targets of clinical relevance.
So far, a few renowned research groups, such as those at the University of Zurich and the University of Tokyo, have successfully developed murine ALPPS models9,10. Creating a standardized murine model under controlled conditions could advance our understanding of the rapid liver regeneration observed after ALPPS procedures.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>5% glucose injection (500 mL)</td>
			<td>Shapuaisi pharma (http://www.zjspas.com/)</td>
			<td>H20003666</td>
			<td>Efficient, cheap,Easy to access</td>
		</tr>
		<tr>
			<td>anaesthesia machine</td>
			<td>RWD (www.rwdls.com)</td>
			<td>R500</td>
			<td>low price and valuable quality. It is suitable for operations with beginners</td>
		</tr>
		<tr>
			<td>C57 BL/6</td>
			<td>The Jackson Lab</td>
			<td>22349-2023</td>
			<td>Stability of strains</td>
		</tr>
		<tr>
			<td>isoflurane</td>
			<td>KCSW (kcsw.szqisoubao.com)</td>
			<td>H19980141</td>
			<td>durable, cost-effective</td>
		</tr>
		<tr>
			<td>meloxicam</td>
			<td>Boehringer-Ingelheim</td>
			<td>H20020217</td>
			<td>Durable and efficient</td>
		</tr>
		<tr>
			<td>microforceps</td>
			<td>maydeal</td>
			<td>60018920</td>
			<td>Durable and efficient</td>
		</tr>
		<tr>
			<td>microinstrument</td>
			<td>CH microsurgical instrument factory</td>
			<td>HC-A804-1</td>
			<td>durable, cost-effective</td>
		</tr>
		<tr>
			<td>sodium lactate ringer</td>
			<td>Shapuaisi pharma (http://www.zjspas.com/)</td>
			<td>H20193277</td>
			<td>Efficient, cheap, easy to access</td>
		</tr>
		<tr>
			<td>suture line</td>
			<td>CH microsurgical instrument factory</td>
			<td>6-0/8-0</td>
			<td>high performance-price ratio</td>
		</tr>
		<tr>
			<td>topical antibodies</td>
			<td>chenxin pharmacology (www.cisen-pharma.com)</td>
			<td>H20020217</td>
			<td>Effectively avoid incision infection</td>
		</tr>
	</tbody>
</table>

a mouse model of the associating liver partition and portal vein ligation for staged hepatectomy procedure aided by microscopy

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative mortality, severely impacting patient outcomes. In a robust hepatic environment, the future liver remnant (FLR) must exceed 25%, and in cases of cirrhosis, this requirement increases to over 40%. The inadequacy of FLR is currently a major obstacle in the progression of hepatic surgery.
Traditional methods to enhance FLR hypertrophy mainly focus on portal vein embolization (PVE), but its effectiveness is considerably limited. In recent years, there have been numerous reports on a novel biphasic hepatectomy method involving hepatic partitioning and portal vein ligation, known as associating liver partition and portal vein ligation for staged hepatectomy (ALPPS). ALPPS surpasses PVE in efficiently and considerably inducing FLR hypertrophy. However, the detailed mechanisms driving ALPPS-facilitated hepatic regeneration are not fully understood. Thus, replicating ALPPS in animal models is crucial to thoroughly investigate the molecular mechanisms of hepatic regeneration, offering valuable theoretical and practical insights.

Author Spotlight: Advancing Liver Regeneration Research through ALPPS Mouse Model

A Mouse Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Procedure Aided by Microscopy

ALPPS can rapidly promote liver regeneration. However, the rather small animal model and the mechanism of the rhythm vision are still insufficient. This model allows for investigating the molecular and cellular pathway involved in liver regeneration of the severe liver injury With was the A of a ARVPS and the PMX mouse models, business studies showed that balance of PMX three and the round two in comparisons regulate haptic was killed under constitution, imposed operative liver regeneration.Single cell sequencing. Single cell attack sequencing. As well as single cell CRISPR has paved the way for the discovery of previously unknown cell types and subtypes in normal and list liver.The ARBs technique is used for patients with liver tumors that cannot be removed through surgery. The ability of the remaining liver to regenerate after the procedure is crucial for the patient's survival. However, the underlying mechanism post AFVBS remaining artery unknown.Based on the previous reports on the rat and the mouse AFVBS GECo models, we constructed a mouse GECo protocol and it was consistent with clinical operation and native limb.

In contrast to the PVL (protocol steps 2,3,4, and 5), the mice that were subjected to the ALPPS (steps 2,3,4,5, and 6) surgical intervention demonstrated a heightened proclivity towards cellular proliferation. Figure 2 delineates the ischemic demarcation of the middle liver lobe following portal vein ligation, along with the visual field during liver transection, which represents a key distinction between ALPPS and PVL. Notably, the levels of inflammatory mediators, including (Tumor Necrosis...

Read this Scientific Article Protocol about A Mouse Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Procedure Aided by Microscopy at JoVE.com

Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) has been shown to result in remarkable liver regeneration after several weeks post first-stage surgery. Animal models that have undergone ALPPS have been developed to explore potential liver regeneration ability and achieve favorable clinical outcomes.

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative ...

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Research

JoVE Journal

Medicine

Associated Liver Partition and Portal Vein Ligation for Staged Hepatectomy for Studying Liver Regeneration in a Mouse Model

To begin, subcutaneously inject one milliliter of lactated Ringer's solution mixed with 5%glucose into the anesthetized mice. Use small animal clippers to shave a two-centimeter area on the abdominal region of the mouse. Then, make a midline incision with a surgical scalpel.Open the incision with a pair of surgical scissors. Use an abdominal retractor to expand the incision, while facilitating the gentle traction of the duodenum and a section of the small intestine with sterile, moist cotton swabs. Cover the structures with a pre-prepared sterile cotton swab to expose the hepatic segment of the portal vein.For portal vein ligation, peel off the peritoneum and intestinal tract. Encircle the main portal vein's branches with slow, deliberate, forward pushing and spreading movements. Identify the right posterior branch, the combined left lateral and median branches, and the caudate branch.Next, to dissect the right posterior branch, locate its prominent, visible branch when traversing the hepatic hilum via the main portal vein. Use moistened cotton balls as a fulling agent to separate the liver's middle lobe from the right posterior lobe. Dissect along the Glisson's capsule.Pull a 1.5-centimeter long silk tie around the portal vein branches. Then, use an 8-0 silk suture to ligate the right posterior branch. For the dissection of the left lateral and median branches, first, dissect any potential interstice between the left lateral lobe and the caudate lobe.Then, introduce a pair of micro forceps into the entrance created. When resistance is felt, adjust the micro forceps slightly to move through the surface of the Glisson's capsule of the right medial lobe. Tie an 8-0 silk suture around the portal vein of the left lateral and median branches.Successful ligation can be observed by the ischemic demarcation within the liver's middle lobe with pallor on the left lobe. Next, dissect the caudal lobe. Fill the potential space between the main portal vein and the caudate lobe with a cotton swab.Peel away the peritoneum of the caudate lobes to visualize the portal vein branches of the lobe. Ligate the caudate branch with a stitch-up suture, ensuring that the needle path adequately wraps the caudate branch. To transect the liver, use an electrocautery pen to create a 0.5-millimeter pre-cut line along the demarcation line in the middle liver lobe.With a pair of micro forceps and the electrocautery pen, cut the liver tissue. Swab the bleeding with cotton. Remove the gallbladder at the end, as it provides a good pulling point through transection.Use a cotton swab to apply controlled pressure for hemostasis. Once transection and cholecystectomy are complete, carefully place the small bowel back into the abdominal cavity. Close the peritoneum and the abdominal wall with 6-0 absorbable sutures.Disinfect the skin's surface, and then subcutaneously inject fentanyl and meloxicam mixed with saline to alleviate postoperative pain. Place the mouse on a thermostatic pad to facilitate awakening. Transfer them into individual cages when they regain consciousness.Subcutaneously inject medetomidine, continuing for 24 hours postoperatively. Mice that were subjected to ALPPS showed a higher proclivity to cellular proliferation. Inflammatory markers of the ALPPS mice, specifically interleukin-6, were higher than those observed in the PVL group.