We would like to acknowledge the support of the Army, Navy NIH, Air Force, VA, and Health Affairs regarding the AFIRM II effort under award CTA05: W81XWH-13-2-0052 and CTA06: W81XWH-13-2-0053. The U.S. Army Medical Research Acquisition Activity, 820 Chandler Street, Fort Detrick MD 21702-5014, is the awarding and administering acquisition office. Opinions, interpretations, conclusions and recommendations are those of the author and are not necessarily endorsed by the Department of Defense. In addition, we would like to acknowledge support from the Department of Defense Congressionally Directed Medical Research Programs (CDMRP), Reconstructive Transplantation Research Program (RTRP), through awards W81XWH-17-1-0280, W81XWH-17-1-0624, W81XWH-17-1-0287, and W81XWH18-1-0795. We would also like to acknowledge the Department of Plastic and Reconstructive Surgery and the Johns Hopkins University School of Medicine. Additionally, we would like to acknowledge the entire veterinary staff, including Melanie Adams, Karen Goss, Haley Smoot, Kayla Schonvisky, and Victoria Manahan.

Hickman Catheter for Long-Term Vascular Access in Swine

<ol>
	<li>Pontes, 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=Incidents+related+to+the+Hickman&#174;+catheter:+identification+of+damages.">Incidents related to the Hickman&#174; catheter: identification of damages.</a> Revista Brasileira de Enfermagem. 71 (4), 1915-1920 (2018).</li><li>Kolikof, J., Peterson, K., Baker, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+Venous+Catheter.">Central Venous Catheter.</a> StatPearls. , (2022).</li><li>Central venous catheters: how, when, why? (Proceedings). DVM 360 Available from: <a target="_blank" href="https://www.dvm360.com/view/central-venous-catheters-how-when-why-proceedings">https://www.dvm360.com/view/central-venous-catheters-how-when-why-proceedings</a> (2011)</li><li>Abrams-Ogg, A. C., 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+use+of+an+implantable+central+venous+(Hickman)+catheter+for+long-term+venous+access+in+dogs+undergoing+bone+marrow+transplantation.">The use of an implantable central venous (Hickman) catheter for long-term venous access in dogs undergoing bone marrow transplantation.</a> Canadian Journal of Veterinary Research. 56 (4), 382-386 (1992).</li><li>Florescu, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+technique+of+placement+of+an+external+jugular+tunneled+hemodialysis+catheter+in+a+large+pig+model.">Surgical technique of placement of an external jugular tunneled hemodialysis catheter in a large pig model.</a> The Journal of Vascular Access. 19 (5), 473-476 (2018).</li><li>. Central Venous Catheter Placement: Modified Seldinger Technique Available from: <a target="_blank" href="https://www.cliniciansbrief.com/article/central-venous-catheter-placement-modified-seldinger-technique">https://www.cliniciansbrief.com/article/central-venous-catheter-placement-modified-seldinger-technique</a> (2015)</li><li>Perondi, 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=Bacterial+colonization+of+non-permanent+central+venous+catheters+in+hemodialysis+dogs.">Bacterial colonization of non-permanent central venous catheters in hemodialysis dogs.</a> Heliyon. 6 (1), e03224 (2020).</li><li>Faulkner, R. T., Czajkowski, W. P., Rayfield, E. J., Hickman, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+for+portal+catheterization+in+rhesus+monkeys+(Macaca+mulatta).">Technique for portal catheterization in rhesus monkeys (Macaca mulatta).</a> American Journal of Veterinary Research. 37 (4), 473-475 (1976).</li><li>Moss, 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=Central+venous+access+devices+for+the+delivery+of+systemic+anticancer+therapy+(CAVA):+a+randomised+controlled+trial.">Central venous access devices for the delivery of systemic anticancer therapy (CAVA): a randomised controlled trial.</a> Lancet. 398 (10298), 403-415 (2021).</li><li>Dai, C., 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+tunneled+and+nontunneled+peripherally+inserted+central+catheter+placement:+A+randomized+controlled+trial.">Effect of tunneled and nontunneled peripherally inserted central catheter placement: A randomized controlled trial.</a> The Journal of Vascular Access. 21 (4), 511-519 (2020).</li><li>Wu, X., 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=Tunneled+peritoneal+catheter+vs+repeated+paracenteses+for+recurrent+ascites:+a+cost-effectiveness+analysis.">Tunneled peritoneal catheter vs repeated paracenteses for recurrent ascites: a cost-effectiveness analysis.</a> Cardiovascular and Interventional Radiology. 45 (7), 972-982 (2022).</li><li>Onwubiko, C., 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=Small+tunneled+central+venous+catheters+as+an+alternative+to+a+standard+hemodialysis+catheter+in+neonatal+patients.">Small tunneled central venous catheters as an alternative to a standard hemodialysis catheter in neonatal patients.</a> Journal of Pediatric Surgery. 56 (12), 2219-2223 (2021).</li><li>da Silva, S. R., Reichembach, M. T., Pontes, L., de Souza, G. d. e. P. E. S. C. M., Kusma, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heparin+solution+in+the+prevention+of+occlusions+in+Hickman&#174;+catheters+a+randomized+clinical+trial.">Heparin solution in the prevention of occlusions in Hickman&#174; catheters a randomized clinical trial.</a> Revista Latino-Americana de Enfermagem. 29, e3385 (2021).</li><li>Landoy, Z., Rotstein, C., Lucey, J., Fitzpatrick, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hickman-Broviac+catheter+use+in+cancer+patients.">Hickman-Broviac catheter use in cancer patients.</a> Journal of Surgical Oncology. 26 (4), 215-218 (1984).</li><li>Bawazir, O. A., Altokhais, T. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hickman+central+venous+catheters+in+children:+open+versus+percutaneous+technique.">Hickman central venous catheters in children: open versus percutaneous technique.</a> Annals of Vascular Surgery. 68, 209-216 (2020).</li><li>Cappello, 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=Central+venous+access+for+haemodialysis+using+the+Hickman+catheter.">Central venous access for haemodialysis using the Hickman catheter.</a> Nephrology Dialysis Transplantation. 4 (11), 988-992 (1989).</li><li>Shastri, L., Kj&#230;rgaard, B., Rees, S. E., Thomsen, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+central+venous+to+arterial+carbon+dioxide+gap+(PCO2+gap)+in+response+to+acute+changes+in+ventilation.">Changes in central venous to arterial carbon dioxide gap (PCO2 gap) in response to acute changes in ventilation.</a> BMJ Open Respiratory Research. 8 (1), e000886 (2021).</li><li>Smith, A. C., Swindle, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+swine+for+the+laboratory.">Preparation of swine for the laboratory.</a> ILAR Journal. 47 (4), 358-363 (2006).</li><li>Swindle, M. M., Makin, A., Herron, A. J., Clubb, F. J., Frazier, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Swine+as+models+in+biomedical+research+and+toxicology+testing.">Swine as models in biomedical research and toxicology testing.</a> Veterinary Pathology. 49 (2), 344-356 (2012).</li><li>Hughes, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Swine+in+cardiovascular+research.">Swine in cardiovascular research.</a> Laboratory Animal Science. 36 (4), 348-350 (1986).</li><li>Svendsen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+minipig+in+toxicology.">The minipig in toxicology.</a> Experimental and Toxicologic Pathology. 57 (5-6), 335-339 (2006).</li><li>Tumbleson, M. E., Schook, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Advances in Swine in Biomedical Research. 2, (1996).</li><li>Jensen-Waern, M., Kruse, R., Lundgren, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+immunosuppressive+medication+for+growing+pigs+in+transplantation+studies.">Oral immunosuppressive medication for growing pigs in transplantation studies.</a> Laboratory Animals. 46 (2), 148-151 (2012).</li><li>Ibrahim, Z., 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+modified+heterotopic+swine+hind+limb+transplant+model+for+translational+vascularized+composite+allotransplantation+(VCA)+research.">A modified heterotopic swine hind limb transplant model for translational vascularized composite allotransplantation (VCA) research.</a> Journal of Visualized Experiments. (80), e50475 (2013).</li><li>Nordstr&#246;m, C. -. H., Jakobsen, R., M&#248;lstr&#248;m, S., Nielsen, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+venous+blood+is+not+drained+via+the+internal+jugular+vein+in+the+pig.">Cerebral venous blood is not drained via the internal jugular vein in the pig.</a> Resuscitation. 162, 437-438 (2021).</li><li>Habib, C. 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=MR+imaging+of+the+yucatan+pig+head+and+neck+vasculature.">MR imaging of the yucatan pig head and neck vasculature.</a> Journal of Magnetic Resonance Imaging. 38 (3), 641-649 (2013).</li><li>Flournoy, W. S., Mani, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+external+jugular+vein+catheterization+in+piglets+using+a+triangulation+technique.">Percutaneous external jugular vein catheterization in piglets using a triangulation technique.</a> The International Journal of Laboratory Animals. 43 (4), 344-349 (2009).</li><li>Kotsougiani, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+angiogenesis+in+porcine+tibial+allotransplantation:+a+new+large+animal+bone+vascularized+composite+allotransplantation+model.">Surgical angiogenesis in porcine tibial allotransplantation: a new large animal bone vascularized composite allotransplantation model.</a> Journal of Visualized Experiments. (126), e55238 (2017).</li><li>Chuang, 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=Comparison+of+external+catheters+with+subcutaneous+vascular+access+ports+for+chronic+vascular+access+in+a+porcine+model.">Comparison of external catheters with subcutaneous vascular access ports for chronic vascular access in a porcine model.</a> Contemporary Topics in Laboratory Animal Science. 44 (2), 24-27 (2005).</li></ol>

None of the authors has a financial interest in any of the products, devices, or drugs mentioned in this manuscript.

While CVCs serve a spectrum of functions in large animal research, current literature lacks a consensus approach for safe and sustainable use in long-term trials over 30 days. This protocol&#39;s stepwise procedure for HC insertion, skin securement, and storage in a handmade pouch has undergone significant adjustments for quality improvement. As such, this protocol presents a technique for HC use that permits efficient and effective intravenous access while ensuring animal welfare and minimizing complications.
<p cla...

The indispensable role of central venous catheters (CVCs) in patient care is owed to their convenience, favorable safety profile, and versatility1. Functions of a CVC include reliable access for total parenteral nutrition, hematopoietic stem cell transplantation, plasmapheresis/apheresis, and efficient fluid, blood, or co-drug administration2. In veterinary medicine, CVCs also minimize animal discomfort via the rapid dilution of irritant drugs and blood sampling without repeated venipuncture3. Despite their broad applications, the use of CVCs in large animal research still presents several considerable challenges4.
Percutaneous CVC placement via a guidewire or introducer catheter can be difficult for non-veterinary researchers, particularly in animals with deep venous structures5. An improper CVC installation technique may result in inadvertent placement into nearby structures, necessitating ultrasound-guided placement or a postprocedure radiography of the positioning6. However, compared to human operating rooms, ultrasounds are not readily available in many large animal research laboratories. Further, long-term use of indwelling catheters can result in line kinking, puncture, infection, or extrication by animals, with the possible disruption of timely treatment, clinical monitoring, and research outcomes4,7. Replacement of the CVC requires additional resources, including material procurement, surgical scheduling, fasting time, and radiographic access. CVC-related complications can therefore create significant technical and financial barriers or a disruption to productive translational research, particularly in swine. Contamination by food or feces, scratching against cage walls, and kicking sites of irritation may compromise a CVC, and the risk of CVC-related complications is amplified by long-term use. Thus, safe and uncomplicated maintenance of a CVC in swine requires careful consideration of CVC choice, placement, securement, protection, sanitation, and surveillance.
The Hickman catheter (HC) used in this protocol is a tunneled CVC with a polyester cuff and one to three lumens, that is commonly used for long-term intravenous access in humans and animals1,4,8,9. The tunneled catheter approach has been associated with lower complication rates and maintenance costs relative to non-tunneled variations10,11,12. The cuff reduces HC extrication by incorporating into the subcutaneous tissues surrounding the skin exit site. The multi-lumen design also enables the separation of medication administration and blood draws, thereby minimizing blood sample contamination and inaccuracy. Despite this, HC use is not without challenges, the most common of which include fracture, migration, occlusion, and infection13,14,15,16. Proper installation and maintenance of a HC are therefore indispensable skills when used in translational research. However, the current literature offers little guidance for best-practices for HC use in swine during long-term trials5,6,17.
The purpose of this study is to outline an optimized approach for HC insertion into the internal jugular vein (IJV), skin securement, and durable protection that minimizes long-term catheter-related complications and discomfort in swine. A discussion of the important considerations for HC use, potential challenges that may be encountered, and modifications that may improve the quality of this approach is included.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
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		<tr>
			<td>#10 blade</td>
			<td>Medline</td>
			<td>MDS15110</td>
			<td></td>
		</tr>
		<tr>
			<td>0.9% Sterile Sodium Chloride</td>
			<td>Baxter&#160;</td>
			<td>2F7123</td>
			<td></td>
		</tr>
		<tr>
			<td>0-0 Coated and Braided Nonabsorbable Suture</td>
			<td>Covidien</td>
			<td>S-196</td>
			<td></td>
		</tr>
		<tr>
			<td>0-0 Synthetic, Monofilament, Nonabsorbable Polypropylene Suture</td>
			<td>Ethicon</td>
			<td>8690H</td>
			<td></td>
		</tr>
		<tr>
			<td>1 inch Medical Tape</td>
			<td>3M</td>
			<td>1548S-1</td>
			<td></td>
		</tr>
		<tr>
			<td>10 USP units/mL Heparin flush</td>
			<td>Becton, Dickinson and Company</td>
			<td>306424</td>
			<td></td>
		</tr>
		<tr>
			<td>3-0 Braided Absorbable Suture</td>
			<td>Covidien</td>
			<td>SL-636 (cutting needle), GL-122 (taper needle)</td>
			<td></td>
		</tr>
		<tr>
			<td>3-0 Monofilament Absorbable Suture</td>
			<td>Covidien</td>
			<td>SM-922 (cutting needle), CM-882 (taper needle)</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 Coated and Braided Non-absorbable Suture Ties</td>
			<td>Ethicon</td>
			<td>A303H</td>
			<td></td>
		</tr>
		<tr>
			<td>70% Ethanol</td>
			<td>Vedco</td>
			<td>VINV-IPA7</td>
			<td></td>
		</tr>
		<tr>
			<td>Adson tissue forceps</td>
			<td>MPM Medical Supply</td>
			<td>132-508</td>
			<td></td>
		</tr>
		<tr>
			<td>Adson-Brown forceps</td>
			<td>MPM Medical Supply</td>
			<td>106-2572</td>
			<td></td>
		</tr>
		<tr>
			<td>Air warming blanket and pad</td>
			<td>3M Bair Hugger</td>
			<td>UPC 00608223595770</td>
			<td></td>
		</tr>
		<tr>
			<td>Backhaus towel clamp</td>
			<td>MPM Medical Supply</td>
			<td>117-5508</td>
			<td></td>
		</tr>
		<tr>
			<td>Brown needle holder</td>
			<td>MPM Medical Supply</td>
			<td>110-1513</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>PAR Pharmaceutical</td>
			<td>3003408B</td>
			<td></td>
		</tr>
		<tr>
			<td>Cefazolin</td>
			<td>Hikma Farmacuetica (Portugal)</td>
			<td>PLB 133-WES/1</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorhexidine</td>
			<td>Vet One</td>
			<td>501027</td>
			<td></td>
		</tr>
		<tr>
			<td>Clave</td>
			<td>Baxter</td>
			<td>7N8399</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Padding</td>
			<td>Medline</td>
			<td>NON6027</td>
			<td></td>
		</tr>
		<tr>
			<td>Debakey forceps</td>
			<td>MPM Medical Supply</td>
			<td>106-5015</td>
			<td></td>
		</tr>
		<tr>
			<td>Elastic Adhesive Bandage Tape</td>
			<td>3M</td>
			<td>XH002016489</td>
			<td></td>
		</tr>
		<tr>
			<td>Halstead mosquito forceps</td>
			<td>MPM Medical Supply</td>
			<td>115-4612</td>
			<td></td>
		</tr>
		<tr>
			<td>Hickman Catheter</td>
			<td>Bard Access Systems</td>
			<td>603710</td>
			<td></td>
		</tr>
		<tr>
			<td>Hickman Catheter Repair Kit, 7Fr, Red and White Connectors</td>
			<td>Bard Access Systems</td>
			<td>0601690 (red), 0601680 (white), 502017</td>
			<td></td>
		</tr>
		<tr>
			<td>Kelly hemostatic forceps</td>
			<td>MPM Medical Supply</td>
			<td>115-7014</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Vet One</td>
			<td>383010-03</td>
			<td></td>
		</tr>
		<tr>
			<td>Lactated Ringers</td>
			<td>Baxter</td>
			<td>2B2324X</td>
			<td></td>
		</tr>
		<tr>
			<td>Maropitant Citrate</td>
			<td>Zoetis</td>
			<td>106</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayo scissors</td>
			<td>MPM Medical Supply</td>
			<td>103-5014</td>
			<td></td>
		</tr>
		<tr>
			<td>Metzenbaum scissors</td>
			<td>MPM Medical Supply</td>
			<td>132-711</td>
			<td></td>
		</tr>
		<tr>
			<td>Pantoprazole</td>
			<td>JH Pharmacy</td>
			<td>NDC 0143-9284-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel blade handle</td>
			<td>Medline</td>
			<td>MDS10801</td>
			<td></td>
		</tr>
		<tr>
			<td>Vein Pick</td>
			<td>SAI infusion technologies</td>
			<td>VP-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary Ophthalmic Ointment</td>
			<td>Dechra</td>
			<td>IS4398</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Vet One</td>
			<td>510004</td>
			<td></td>
		</tr>
	</tbody>
</table>

hickman catheter use for long-term vascular access in a preclinical swine model

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood monitoring and reliable intravenous fluid and drug administration. Specifically, the tunneled multi-lumen Hickman catheter (HC) is commonly used in swine models due to its lower extrication and complication rates. Despite fewer complications relative to other CVCs, HC-related morbidity presents a significant challenge, as it can significantly delay or otherwise negatively impact ongoing studies. The proper insertion and maintenance of HCs is paramount in preventing these complications, but there is no consensus on best practices. The purpose of this protocol is to comprehensively describe an approach for the insertion and maintenance of a tunneled HC in swine that mitigates HC-related complications and morbidity. The use of these techniques in &gt;100 swine has resulted in complication-free patent lines up to 8 months and no catheter-related mortality or infection of the ventral surgical site. This protocol offers a method to optimize the lifespan of the HC and guidance for approaching issues during use.

Over 100 swine have undergone successful HC insertion in our lab. The HC can be safely and correctly placed and secured in under 1 h with a surgeon, assistant, circulator, and anesthetist. The catheter pouch takes roughly 15-20 min to make. The technique is straightforward and easy to teach and has been performed by veterinarians, surgical residents, and medical students following supervised instructions.
HCs have remained in place without complications or revision for up to 8 months. In a rec...

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Author Spotlight: Hickman Catheter Use for Long-Term Vascular Access in a Preclinical Swine Model  

A reliable and reproducible approach for the insertion and maintenance of a tunneled Hickman catheter for long-term vascular access in swine is described. Placement of a central venous catheter allows for convenient daily sampling of whole blood from awake animals and intravenous administration of medication and fluids.

Hickman Catheter Use for Long-Term Vascular Access in a Preclinical Swine Model

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood ...

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Medicine

3.7K Views.  Johns Hopkins University School of Medicine. A reliable and reproducible approach for the insertion and maintenance of a tunneled Hickman catheter for long-term vascular access in swine is described. Placement of a central venous catheter allows for convenient daily sampling of whole blood from awake animals and intravenous administration of medication and fluids. 

Video: Author Spotlight: Hickman Catheter Use for Long-Term Vascular Access in a Preclinical Swine Model  

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

A Swine Model of Neonatal Asphyxia

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, perfusion deficit, hypoxic-ischemic encephalopathy, pulmonary hypertension, vasculopathic enterocolitis, renal failure and thrombo-embolic complications. Animal models are developed to help us understand the patho-physiology and pharmacology of neonatal asphyxia. In comparison to rodents and newborn lambs, the newborn piglet has been proven to be a valuable model. The newborn piglet has several advantages including similar development as that of 36-38 weeks human fetus with comparable body systems, large body size (&tilde;1.5-2 kg at birth) that allows the instrumentation and monitoring of the animal and controls the confounding variables of hypoxia and hemodynamic derangements.
We here describe an experimental protocol to simulate neonatal asphyxia and allow us to examine the systemic and regional hemodynamic changes during the asphyxiating and reoxygenation process as well as the respective effects of interventions. Further, the model has the advantage of studying multi-organ failure or dysfunction simultaneously and the interaction with various body systems. The experimental model is a non-survival procedure that involves the surgical instrumentation of newborn piglets (1-3 day-old and 1.5-2.5 kg weight, mixed breed) to allow the establishment of mechanical ventilation, vascular (arterial and central venous) access and the placement of catheters and flow probes (Transonic Inc.) for the continuously monitoring of intra-vascular pressure and blood flow across different arteries including main pulmonary, common carotid, superior mesenteric and left renal arteries. Using these surgically instrumented piglets, after stabilization for 30-60 minutes as defined by Z&lt;10% variation in hemodynamic parameters and normal blood gases, we commence an experimental protocol of severe hypoxemia which is induced via normocapnic alveolar hypoxia. The piglet is ventilated with 10-15% oxygen by increasing the inhaled concentration of nitrogen gas for 2h, aiming for arterial oxygen saturations of 30-40%. This degree of hypoxemia will produce clinical asphyxia with severe metabolic acidosis, systemic hypotension and cardiogenic shock with hypoperfusion to vital organs. The hypoxia is followed by reoxygenation with 100% oxygen for 0.5h and then 21% oxygen for 3.5h. Pharmacologic interventions can be introduced in due course and their effects investigated in a blinded, block-randomized fashion.

Large animal models have good translational values in the examination of physiology and pharmacology of neonatal asphyxia. Using newborn piglets, we develop an experimental protocol to simulate neonatal asphyxia which has advantages of studying the systemic and regional hemodynamics, oxygen transport with biochemical and pathologic pathways and correlations.

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, ...

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

Severe Burn Injury in a Swine Model for Clinical Dressing Assessment

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological pathways impair the inflammation response, resulting in delayed wound healing. Impaired wound healing frequently occurs in patients with diabetes leading to unfavorable outcomes such as amputation. Hence, dressings having beneficial effect in promoting burn wound repair are needed. However, studies on burn wound treatment are limited due to lack of proper animal models. Our previous study demonstrated wound-healing performance in rat and swine models using a minimally invasive surgical technique. This study aimed to demonstrate a swine model of severe burn injury that eliminates wound contraction and more closely approximates the human processes of re-epithelialization and new tissue formation. This protocol provides a detailed procedure for creating consistent burn wounds and examining the wound-healing performance under the treatment of an experimental dressing in a swine model. Six burn wounds were created symmetrically on the dorsum, which were covered with a clinical dressing composed of four layers: an inner contact layer of experimental materials, an inner intermediate layer of waterproof film, an outer intermediate layer of gauze, and an outer layer of adhesive plaster. Upon the completion of experiments, wound closure, wound area, and Vancouver Scar Scale score were examined. The samples of skin resected from each animal post-sacrifice were histologically prepared and stained using hematoxylin and eosin staining. Antibacterial activity of each dressing in the context of wound healing was also examined. The application of the clinical dressing to the wounds in swine model mimics the biological processes of human wound healing with respect to the processes of epithelialization, cellular proliferation, and angiogenesis. Therefore, this swine model provides an easy-to-learn, cost-effective, and robust method to assess the effect of clinical dressings in severe burn injury.

To closely mimic the mode of burn injuries requires the interplay between clinical observation and studies in animal models. In this study, a swine model of severe burn injury was established to assess an experimental dressing in physiological and pathophysiological settings.

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

Analyzing Long-Term Electrocardiography Recordings to Detect Arrhythmias in Mice

Arrhythmias are common, affecting millions of patients worldwide. Current treatment strategies are associated with significant side effects and remain ineffective in many patients. To improve patient care, novel and innovative therapeutic concepts causally targeting arrhythmia mechanisms are needed. To study the complex pathophysiology of arrhythmias, suitable animal models are necessary, and mice have been proven to be ideal model species to evaluate the genetic impact on arrhythmias, to investigate fundamental molecular and cellular mechanisms, and to identify potential therapeutic targets.
Implantable telemetry devices are among the most powerful tools available to study electrophysiology in mice, allowing continuous ECG recording over a period of several months in freely moving, awake mice. However, due to the huge number of data points (&gt;1 million QRS complexes per day), analysis of telemetry data remains challenging. This article describes a step-by-step approach to analyze ECGs and to detect arrhythmias in long-term telemetry recordings using the software, Ponemah, with its analysis modules, ECG Pro and Data Insights, developed by Data Sciences International (DSI). To analyze basic ECG parameters, such as heart rate, P wave duration, PR interval, QRS interval, or QT duration, an automated attribute analysis was performed using Ponemah to identify P, Q, and T waves within individually adjusted windows around detected R waves. 
Results were then manually reviewed, allowing adjustment of individual annotations. The output from the attribute-based analysis and the pattern recognition analysis was then used by the Data Insights module to detect arrhythmias. This module allows an automatic screening for individually defined arrhythmias within the recording, followed by a manual review of suspected arrhythmia episodes. The article briefly discusses challenges in recording and detecting ECG signals, suggests strategies to improve data quality, and provides representative recordings of arrhythmias detected in mice using the approach described above.

Here we present a step-by-step protocol for a semiautomated approach to analyze murine long-term electrocardiography (ECG) data for basic ECG parameters and common arrhythmias. Data are obtained by implantable telemetry transmitters in living and awake mice and analyzed using Ponemah and its analysis modules.

Arrhythmias are common, affecting millions of patients worldwide. Current treatment strategies are associated with significant side effects and remain ...

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Technique for Obtaining Mesenchymal Stem Cell from Adipose Tissue and Stromal Vascular Fraction Characterization in Long-Term Cryopreservation

Human mesenchymal stem cells derived from adipose tissue have become increasingly attractive as they show appropriate features and are an accessible source for regenerative clinical applications. Different protocols have been used to obtain adipose-derived stem cells. This article describes different steps of an improved time-saving protocol to obtain a more significant amount of ADSC, showing how to cryopreserve and thaw ADSC to obtain viable cells for culture expansion. One hundred milliliters of lipoaspirate were collected, using a 26 cm three-hole and 3 mm caliber syringe liposuction, from the abdominal area of nine patients who subsequently underwent elective abdominoplasty. The stem cells isolation was carried out with a series of washes with Dulbecco&#39;s Phosphate Buffered Saline (DPBS) solution supplemented with calcium and the use of collagenase. Stromal Vascular Fraction (SVF) cells were cryopreserved, and their viability was checked by immunophenotyping. The SVF cellular yield was 15.7 x 105 cells/mL, ranging between 6.1-26.2 cells/mL. Adherent SVF cells reached confluence after an average of 7.5 (&#177;4.5) days, with an average cellular yield of 12.3 (&#177; 5.7) x 105 cells/mL. The viability of thawed SVF after 8 months, 1 year, and 2 years ranged between 23.06%-72.34% with an average of 47.7% (&#177;24.64) with the lowest viability correlating with cases of two-year freezing. The use of DPBS solution supplemented with calcium and bag resting times for fat precipitation with a shorter time of collagenase digestion resulted in an increased stem cell final cellular yield. The detailed procedure for obtaining high yields of viable stem cells was more efficient regarding time and cellular yield than the techniques from previous studies. Even after a long period of cryopreservation, viable ADSC cells were found in the SVF.

The present protocol describes an improved methodology for ADSC isolation resulting in a tremendous cellular yield with time gain compared to the literature. This study also provides a straightforward method for obtaining a relatively large number of viable cells after long-term cryopreservation.

Human mesenchymal stem cells derived from adipose tissue have become increasingly attractive as they show appropriate features and are an accessible ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

Preparation of Human Myocardial Tissue for Long-Term Cultivation

Cardiomyocyte cultivation has seen a vast number of developments, ranging from two-dimensional (2D) cell cultivation to iPSC derived organoids. In 2019, an ex vivo way to cultivate myocardial slices obtained from human heart samples was demonstrated, while approaching in vivo condition of myocardial contraction. These samples originate mostly from heart transplantations or left-ventricular assist device placements. Using a vibratome and a specially developed cultivation system, 300 &#181;m thick slices are placed between a fixed and a spring wire, allowing for stable and reproducible cultivation for several weeks. During cultivation, the slices are continuously stimulated according to individual settings. Contractions can be displayed and recorded in real-time, and pharmacological agents can be readily applied. User-defined stimulation protocols can be scheduled and performed to assess vital contraction parameters like post-pause-potentiation, stimulation threshold, force-frequency relation, and refractory period. Furthermore, the system enables a variable pre- and afterload setting for a more physiological cultivation.
Here, we present a step-by-step guide on how to generate a successful long-term cultivation of human left ventricular myocardial slices, using a commercial biomimetic cultivation solution.

We present a protocol for ex vivo cultivation of human ventricular myocardial tissue. It allows for detailed analysis of contraction force and kinetics, as well as the application of pre- and afterload to mimic the in vivo physiological environment more closely.

Cardiomyocyte cultivation has seen a vast number of developments, ranging from two-dimensional (2D) cell cultivation to iPSC derived organoids. In 2019, ...