Name: use of a central venous line for fluids, drugs and nutrient administration in a mouse model of critical illness
Uploaded: 2017-05-02T14:45:00
Description: Watch this Scientific Journal Video about Use of a Central Venous Line for Fluids, Drugs and Nutrient Administration in a Mouse Model of Critical Illness at JoVE.com

GVdB, via the University of Leuven (KU Leuven), receives long-term structural research support from the Methusalem Program funded by the Flemish Government (METH08/07) and holds a European Research Council Advanced Grant AdvG-2012-321670 from the Ideas Program of the European Union seventh framework program. SET received a Research Foundation-Flanders (FWO) Research Assistant Fellowship.

The protocol was approved by the University of Leuven Ethical Review Board for Animal Research.
1. Preparation of the Venous Line
<ol>
	<li>Prepare the tip of the venous catheter by quickly immersing the middle part of 60 cm microrenathane (MRE) tubing in hot (&#62; 220 &#176;C) sesame oil. Subsequently stretch the middle part of the tube to produce a narrow diameter (outer diameter (OD) &#60; 0.5 mm) by gently moving the ends of the tubing away from each other.</li>
	<li>Use a scalpel blade...

<ol>
	<li>Angus, D. C., van der Poll, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Severe+sepsis+and+septic+shock.">Severe sepsis and septic shock.</a> N Engl J Med. 369 (9), 840-851 (2013).</li><li>Singer, 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=The+Third+International+Consensus+Definitions+for+Sepsis+and+Septic+Shock+(Sepsis-3).">The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3).</a> JAMA. 315 (8), 801-810 (2016).</li><li>Remick, D. G., Newcomb, D. E., Bolgos, G. L., Call, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+mortality+and+inflammatory+response+of+two+models+of+sepsis:+lipopolysaccharide+vs.+cecal+ligation+and+puncture.">Comparison of the mortality and inflammatory response of two models of sepsis: lipopolysaccharide vs. cecal ligation and puncture.</a> Shock. 13 (2), 110-116 (2000).</li><li>Copeland, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+inflammatory+response+to+endotoxin+in+mice+and+humans.">Acute inflammatory response to endotoxin in mice and humans.</a> Clin Diagn Lab Immunol. 12 (1), 60-67 (2005).</li><li>Seok, 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=Genomic+responses+in+mouse+models+poorly+mimic+human+inflammatory+diseases.">Genomic responses in mouse models poorly mimic human inflammatory diseases.</a> Proc Natl Acad Sci U S A. 110 (9), 3507-3512 (2013).</li><li>Rittirsch, D., Huber-Lang, M. S., Flierl, M. A., Ward, 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=Immunodesign+of+experimental+sepsis+by+cecal+ligation+and+puncture.">Immunodesign of experimental sepsis by cecal ligation and puncture.</a> Nat Protoc. 4 (1), 31-36 (2009).</li><li>Zolfaghari, P. S., Pinto, B. B., Dyson, A., Singer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+metabolic+phenotype+of+rodent+sepsis:+cause+for+concern.">The metabolic phenotype of rodent sepsis: cause for concern.</a> Intensive Care Med Exp. 1 (1), 25 (2013).</li><li>Radermacher, P., Haouzi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+is+not+a+rat+is+not+a+man:+species-specific+metabolic+responses+to+sepsis+-+a+nail+in+the+coffin+of+murine+models+for+critical+care+research.">A mouse is not a rat is not a man: species-specific metabolic responses to sepsis - a nail in the coffin of murine models for critical care research.</a> Intensive Care Med Exp. 1 (1), 26 (2013).</li><li>Dyson, A., Singer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+sepsis:+why+does+preclinical+efficacy+fail+to+translate+to+the+clinical+setting.">Animal models of sepsis: why does preclinical efficacy fail to translate to the clinical setting.</a> Crit Care Med. 37 (1 Suppl), S30-S37 (2009).</li><li>Osuchowski, M. F., 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=Abandon+the+mouse+research+ship?+Not+just+yet.">Abandon the mouse research ship? Not just yet.</a> Shock. 41 (6), 463-475 (2014).</li><li>Langford DJ1, B. A., Chanda, M. L., Clarke, S. E., Drummond, T. E., Echols, S., Glick, S., Ingrao, J., Klassen-Ross, T., Lacroix-Fralish, M. L., Matsumiya, L., Sorge, R. E., Sotocinal, S. G., Tabaka, J. M., Wong, D., van den Maagdenberg, A. M., Ferrari, M. D., Craig, K. D., Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+facial+expressions+of+pain+in+the+laboratory+mouse.">Coding of facial expressions of pain in the laboratory mouse.</a> Nat Methods. 7, (2010).</li><li>Hollenberg, S. 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=Characterization+of+a+hyperdynamic+murine+model+of+resuscitated+sepsis+using+echocardiography.">Characterization of a hyperdynamic murine model of resuscitated sepsis using echocardiography.</a> Am J Respir Crit Care Med. 164 (5), 891-895 (2001).</li><li>Heuer, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cecal+ligation+and+puncture+with+total+parenteral+nutrition:+a+clinically+relevant+model+of+the+metabolic,+hormonal,+and+inflammatory+dysfunction+associated+with+critical+illness.">Cecal ligation and puncture with total parenteral nutrition: a clinically relevant model of the metabolic, hormonal, and inflammatory dysfunction associated with critical illness.</a> J Surg Res. 121 (2), 178-186 (2004).</li><li>Conti, M., Merlani, P., Ricou, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prognosis+and+quality+of+life+of+elderly+patients+after+intensive+care.">Prognosis and quality of life of elderly patients after intensive care.</a> Swiss Med Wkly. 142, (2012).</li></ol>

We developed a more clinically relevant mouse model of critical illness, by combining the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration. This experimental setup is reproducible, allows to study the prolonged phase of critical illness and results in a stable mortality rate, hereby mimicking the human clinical situation1.
Cecal ligation and puncture has been extensively recognized as...

Critical illness is a disease state in which the function of one or more organ systems is hampered to the extent that the patient will die, unless intensive medical support is administered. Whereas the initial cause for admission to the intensive care unit (ICU) can vary, ranging from trauma, complicated surgery, burns, disease exacerbations to sepsis, all critically ill patients suffer from cellular damage, caused by hypoperfusion, hypoxia and excessive inflammation among others, which leads to organ failure. Most patients survive their acute insult, but an important fraction of the patients do not immediately recover and need prolonged intensive care. They are at high risk of death due to non-resolving multiple organ failure. Furthermore, the prolonged phase of critical illness is hallmarked by profound muscle weakness and endocrine and metabolic change,&#160; of which the pathogenesis is currently incompletely understood.
Several rodent models are being used in the critical care research setting. The two mostly used models are the exogenous administration of lipopolysaccharide (LPS) and cecal ligation and puncture (CLP). Both models are developed to mimic the acute phase of sepsis, defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection, and one of the primary reasons for admission to the ICU worldwide1,2. The LPS model has several disadvantages as it only transiently affects the release of cytokines and the hemodynamic status of the animal3. Unlike humans, rodents are also particularly resistant to endotoxin and the use of &#39;high doses&#39; of endotoxin is necessary to produce hypotension and mortality, hereby further raising concerns of the validity of this method4,5. The other model, cecal ligation and puncture model (CLP) features a ligation of a portion of the cecum followed by a needle puncture through-and-through. This procedure causes a polymicrobial abdominal infection with tissue damage, followed by a translocation of the bacteria into the blood compartment. This will trigger a systemic inflammatory response and the development of sepsis. The CLP model has been extensively recognized as an animal model of acute critical illness that reproduces the main features of sepsis: hyperinflammation, vasodilation, hypotension and increased cardiac output6,7. However, this model does not allow study of non-resolving multiple organ failure, muscle wasting, and endocrine and metabolic changes, which are typical for the prolonged phase of critical illness. Furthermore, recently the validity of mouse models for critical illness have been questioned, since findings from mouse models cannot always be translated to the human setting8,9,10. A possible explanation might be that the supportive care that is provided to critically ill human patients differs substantially from the care that is provided to critically ill mice.
Therefore, to resemble the human setting more closely and to allow investigation of the prolonged phase of critical illness, we developed a mouse model that mimics the acute intensive care as given to humans, such as extensive intravenous fluid administration and antibiotic treatment, and which allows to administer supportive care in order to survive the prolonged phase of critical illness, such as nutritional support. For this purpose, we adapted the mouse model of CLP-induced sepsis, being the golden standard for sepsis, and placed a central venous line which enables the administration of fluids, nutrition and drugs.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Buprenorphine</td>
			<td>Ecuphar</td>
			<td></td>
			<td>Vetergesic 0,3mg/ml</td>
		</tr>
		<tr>
			<td>C57BL/6&#160;</td>
			<td>Janvier labs</td>
			<td>C57BL/6JRj</td>
			<td></td>
		</tr>
		<tr>
			<td>colloids</td>
			<td>Fresenius Kabi</td>
			<td></td>
			<td>Volulyte 6%</td>
		</tr>
		<tr>
			<td>crystalloids</td>
			<td>Baxter</td>
			<td></td>
			<td>Plasmalyte a viaflo</td>
		</tr>
		<tr>
			<td>Ethilon 3.0</td>
			<td>Ethicon</td>
			<td>F3211</td>
			<td></td>
		</tr>
		<tr>
			<td>Imipenem</td>
			<td>MSD&#160;</td>
			<td></td>
			<td>Tienam 500mg powder for injection fluid</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Eurovet</td>
			<td></td>
			<td>Iso-vet</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Eurovet</td>
			<td></td>
			<td>Nimatek 100mg/ml</td>
		</tr>
		<tr>
			<td>LocTite Super glue3 all plastics&#160;</td>
			<td>Rectavit</td>
			<td>119818</td>
			<td></td>
		</tr>
		<tr>
			<td>Mersilk 3.0</td>
			<td>Ethicon</td>
			<td>L192</td>
			<td></td>
		</tr>
		<tr>
			<td>Mersilk 5.0</td>
			<td>Ethicon</td>
			<td>F682</td>
			<td></td>
		</tr>
		<tr>
			<td>Microrenathane .025 O.D.&#34; x .012 I.D.&#34;</td>
			<td>Bioseb</td>
			<td>MRE-025</td>
			<td></td>
		</tr>
		<tr>
			<td>olimel N7E</td>
			<td>Baxter</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PE10 - Polyethylene .011&#34; x .024&#34;&#160;</td>
			<td>Instech Solomon</td>
			<td>BTPE-10</td>
			<td></td>
		</tr>
		<tr>
			<td>PE-50 tubing .023&#34;x.038&#34;</td>
			<td>Instech Solomon</td>
			<td>BTPE-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Rodent Swivel 20 G</td>
			<td>Bioseb</td>
			<td>RS-20G&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Ropivaca&#239;ne</td>
			<td>Astrazenica</td>
			<td></td>
			<td>Naropin 2mg/ml</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>VMD</td>
			<td></td>
			<td>Xylazine hydrochloride 2%</td>
		</tr>
	</tbody>
</table>

use of a central venous line for fluids, drugs and nutrient administration in a mouse model of critical illness

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting. Critically ill patients require intensive medical support in order to survive. While the majority of patients will recover within a few days, about a quarter of the patients need prolonged intensive care and are at high risk of dying from non-resolving multiple organ failure. Furthermore, the prolonged phase of critical illness is hallmarked by profound muscle weakness, and endocrine and metabolic changes, of which the pathogenesis is currently incompletely understood. The most widely used animal model in critical care research is the cecal ligation and puncture model to induce sepsis. This is a very reproducible model, with acute inflammatory and hemodynamic changes similar to human sepsis, which is designed to study the acute phase of critical illness. However, this model is hallmarked by a high lethality, which is different from the clinical human situation, and is not developed to study the prolonged phase of critical illness. Therefore, we adapted the technique by placing a central venous catheter in the jugular vein allowing us to administer clinically relevant supportive care, to better mimic the human clinical situation of critical illness. This mouse model requires an extensive surgical procedure and daily intensive care of the animals, but it results in a relevant model of the acute and prolonged phase of critical illness.

C57BL/6 mice were made critically ill as described above. We performed two experiments to assess post-CLP survival until two time points: survival until day 5 (n = 15) and survival until day 7 (n = 22) post-CLP. Survival curves of the two experiments were not significantly different (compared until day 5), indicating the reproducibility of the experimental setup. Non&#8211;surviving animals were found death or euthanized due to reaching human endpoints.&#160;&#160;Ligation of 50% of the c...

Watch this Scientific Journal Video about Use of a Central Venous Line for Fluids, Drugs and Nutrient Administration in a Mouse Model of Critical Illness at JoVE.com

Use of a Central Venous Line for Fluids, Drugs and Nutrient Administration in a Mouse Model of Critical Illness

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting.

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce ...

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