Name: lung rapid recovery procurement combined with abdominal normothermic regional perfusion in controlled donation after circulatory death
Uploaded: 2022-08-15T14:00:00
Description: Watch this Scientific Journal Video about Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death at JoVE.com

The authors acknowledge all the members involved in the Lung Transplant Program at University Hospital Marqu&#233;s de Valdecilla.

These interventions are undertaken at the bedside in the intensive care unit (ICU). This protocol follows the guidelines of the University Hospital Marqu&#233;s de Valdecilla ethics committee and is in accordance with the Spanish legal framework regarding donation procedures. Informed consent was obtained from next of kin for video recording of the procedures for research. cDCD is considered in patients with catastrophic brain damage or a terminal heart or a neurodegenerative disease for whom the decision to withdraw lif...

Though the use of simultaneous lung cold perfusion with ARNP in cDCD was first published in 2014, very few experiences have been described for this25,26,29. Moreover, the utilization of cDCD lungs, regardless of the technique used, remains low in most countries.
The critical steps within this protocol are the use of premortem interventions; a specific methodology to ensure coronary and cerebral perfus...

Donation after circulatory death (DCD) started in Spain with uncontrolled donors. In 1996, the first national consensus document on DCD was published as a guide for the practice of uncontrolled donation after circulatory death1 (uDCD), also setting a moratorium on controlled donation after circulatory death (cDCD). In 2012, a new consensus emerged establishing the basis and the legislative framework for the practice of both uDCD and cDCD2. Currently, Spain is one of the most active countries in DCD, reaching the highest rate of donors after circulatory death in the world3. This type of donor represented nearly 35% of the total donors in 2021 in the country, with a marked decrease in uDCD and donors being exclusively cDCD4.
Organ procurement in cDCD is commonly performed using the super-rapid recovery technique5. After the declaration of death and when the non-touch period has elapsed, a rapid sternotomy and laparotomy is performed. The abdominal aorta and pulmonary artery are cannulated and flushed with cold perfusion solutions to preserve the abdominal and thoracic organs, plus topical cooling is conducted before retrieval6. In this situation, cDCD is characterized by the unpredictable consequences of warm ischemia,&#160;after the withdrawal of life-sustaining therapy. The ischemic damage during this period of agonic hypotension and progressive hypoxia, followed by the non-touch period after cardiac arrest, is further exacerbated by the later period of cold ischemia7. This combination of warm and cold ischemia seems to be detrimental, especially to abdominal grafts8,9,10,&#160;generating more reluctance among professionals in the use of these organs from cDCD donors.
To minimize these risks, an in situ preservation model, based on previous experiences from Spanish teams working in uCDC11,&#160;has been developed with growing interest. The use of extracorporeal membrane oxygenation (ECMO) systems to restore blood flow after death and before graft recovery can reverse the metabolic deviations resulting from ischemia and restore cellular physiology12. Abdominal normothermic regional perfusion (ANRP) can improve the quality of ischemic-damaged organs in cDCD13. Organ function can be assessed and improved, allowing a better selection of abdominal grafts for transplantation.
Recent international multicenter experiences provide evidence that ANRP versus the rapid recovery (RR) technique helps overcome traditional limitations in cDCD, reducing rates of post-transplant biliary complications, facilitating successful transplantation of older livers, and improving liver graft survival14,15. In kidneys, it seems to improve short-term outcomes with a lower delayed graft function and higher 1 year graft survival rates16. With this evidence, ANRP in cDCD has gained advantages over the rapid recovery technique for abdominal graft procurement and is now applied in several European countries and other parts of the world17,18.
The use of lungs from cDCD donors, however, was promptly adopted worldwide. A lung functional warm ischemic time of up to 60 min does not seem to affect survival19. In the last decade, several centers and multi-institutional experiences have reported outcomes after lung transplant from cDCD comparable to those from DBD20,21. The RR technique is the routine method for lung procurement: lungs are cooled topically and removed after being flushed with cold preservation solution22.
The first experiences combining ANRP and RR of lungs in cDCD were reported by two United Kingdom groups23,24. Years later, a variation of this technique adding premortem interventions was published25. Results present this dual procurement technique as safe and effective for both abdominal and thoracic grafts26. Obviously, the donation procedure becomes more complex. It requires technological and human resources, sufficient organizational capabilities, and has a higher economic cost. All of this may discourage professionals from starting a program. The aim of this study is to present a protocol especially focused on premortem interventions, cannulation, and aortic occlusion balloon placement, with tips and tricks learned from experience, and comment the different technical details to consider during lung retrieval when ARNP is used. At present, in the Center, cDCD donors have become the main source of grafts for thoracic and abdominal transplantation.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Vial 5 mL Heparin 1000 UI/mL</td>
			<td>ROVI</td>
			<td></td>
			<td>For donor heparinization</td>
		</tr>
		<tr>
			<td>ECMO KIT (MATERIALS FOR CANNULATION)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Artery pressure lines</td>
			<td>BEXEN MEDICAL</td>
			<td>137.15</td>
			<td>Artery pressure line por radial artery and femoral cannula</td>
		</tr>
		<tr>
			<td>Bandage scissors</td>
			<td>SURGIMEDIC</td>
			<td>BC-881R</td>
			<td>Shear to cut ECMO lines</td>
		</tr>
		<tr>
			<td>Bio-medicus Venous cannula 21 Fr (7.0 mm) x 27.5 in (69.9 cm)</td>
			<td>MEDTRONIC</td>
			<td>96670-121</td>
			<td>Venous cannula</td>
		</tr>
		<tr>
			<td>Clhorhexidine solution 2%</td>
			<td></td>
			<td></td>
			<td>Disinfectant solution</td>
		</tr>
		<tr>
			<td>ECMO device Maquet Rotaflow</td>
			<td>Maquet, Rasttat, Germany</td>
			<td></td>
			<td>ECMO system</td>
		</tr>
		<tr>
			<td>Electrocautery handle</td>
			<td>DEXTRO</td>
			<td>SW12200</td>
			<td></td>
		</tr>
		<tr>
			<td>EndoReturn Arterial Cannula Kit&#160; 21-23F</td>
			<td>Edwards Lifesciences</td>
			<td>ER21B, ER23B</td>
			<td>Arterial cannula with a doble lumen to ECMO connection and to introduce aortic oclussion balloon</td>
		</tr>
		<tr>
			<td>Ethicon LigaClip med/short 20 titanium medium</td>
			<td>ETHICON</td>
			<td>MCS30</td>
			<td>Ligaclips for control bleeding during groin dissection</td>
		</tr>
		<tr>
			<td>Ethicon LigaClip med/short 20 titanium small</td>
			<td>ETHICON</td>
			<td>MCS20</td>
			<td>Ligaclips for control bleeding during groin dissection</td>
		</tr>
		<tr>
			<td>Insertion Kit Bio-medicus 180cm</td>
			<td>MEDTRONIC</td>
			<td>96551</td>
			<td>Insertion Kit for ECMO cannulas, with catheter, metal wire guide and dilators</td>
		</tr>
		<tr>
			<td>Irrigation pear</td>
			<td>MEDLINE</td>
			<td>DYNDE 20125</td>
			<td>Pear to be filled with saline and purge ECMO lines at the site of connection with cannulas</td>
		</tr>
		<tr>
			<td>Luer cone syringe 50cc</td>
			<td>CARDIONATUR</td>
			<td>60ML</td>
			<td>Syringe filled with saline to fill occlusion balloon</td>
		</tr>
		<tr>
			<td>Mersilk no 1, LR-60 CONV , 75 cm</td>
			<td>ETHICON</td>
			<td>W562H</td>
			<td>Silk curved suture for ECMO cannulas fixation</td>
		</tr>
		<tr>
			<td>Prolene 4/0</td>
			<td>ETHICON</td>
			<td>W8355</td>
			<td>polypropylene suture for purse string in femoral vessels or vascular suture</td>
		</tr>
		<tr>
			<td>Prolene 5/0 , 60 cm</td>
			<td>ETHICON</td>
			<td>8325</td>
			<td>polypropylene suture for vascular suture</td>
		</tr>
		<tr>
			<td>Prolene 5/0, 90 cm</td>
			<td>ETHICON</td>
			<td>8720</td>
			<td>polypropylene suture for vascular suture</td>
		</tr>
		<tr>
			<td>Reliant Stent Graft Balloon Catheter&#160;12F</td>
			<td>Medtronic, Ireland</td>
			<td>AB46</td>
			<td>Aortic occlusion balloon introduced through femoral artery. It is used as an endoclamp</td>
		</tr>
		<tr>
			<td>Scalpel blade no 11</td>
			<td>INTRAVEN</td>
			<td>150011</td>
			<td></td>
		</tr>
		<tr>
			<td>Scapel blade no 23</td>
			<td>INTRAVEN</td>
			<td>150023</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicone tube</td>
			<td>IBERHOSPITEX</td>
			<td>0027224-P</td>
			<td>Silicone tube to connect suction system</td>
		</tr>
		<tr>
			<td>Sofsilk braided silk no 1 strands</td>
			<td>COVIDIEN</td>
			<td>L-12</td>
			<td>Silk strand for ligation or bleeding control</td>
		</tr>
		<tr>
			<td>Sofsilk braided silk no 3 strands</td>
			<td>COVIDIEN</td>
			<td>L-115</td>
			<td>Silk strand for ligation or bleeding control</td>
		</tr>
		<tr>
			<td>straight connector 3/8&#34;x3/8&#34; with Luer lock</td>
			<td>ANDOCOR</td>
			<td>04CS0022</td>
			<td>Piece to connect arterial cannula with ECMO line and the three way stop-cock for pressure line and blood sampling</td>
		</tr>
		<tr>
			<td>Surgical pads pack</td>
			<td>TEXPOL</td>
			<td>146500</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical stapler</td>
			<td>COVIDIEN</td>
			<td>8886803712</td>
			<td>Stapler to close surgical wound</td>
		</tr>
		<tr>
			<td>Three-way stopcock</td>
			<td>BD CONNECTA</td>
			<td>394501</td>
			<td>Three way stop-cock to connect farterial cannula with pressure line</td>
		</tr>
		<tr>
			<td>Vessel loop large</td>
			<td>MEDLINE</td>
			<td>VLMAXR</td>
			<td>Vascular loop to embrace femoral artery and vein for bleeding control.</td>
		</tr>
		<tr>
			<td>Vessel loop small</td>
			<td>MEDLINE</td>
			<td>VLMINR</td>
			<td>Vascular loop to embrace femoral artery and vein for bleeding control.</td>
		</tr>
		<tr>
			<td>Yankauer suction terminal 50 V</td>
			<td>DEXTROMEDICA</td>
			<td>349701</td>
			<td>Suction terminal for suction while surgical dissection</td>
		</tr>
		<tr>
			<td>SURGICAL TOOLS FOR CANNULATION</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Adson retractor 20 cm adn 33 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Aortic clamp</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Boyd Scissors 18 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissection forceps without jaws 21 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Farabeuf retractor small</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mayo scissors straight 14 cm and 16 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Metzembaum scissors 18 cm, 20 cm and 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito forceps straigth and curved</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder 18 cm and 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Russ dissection forceps 15 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel handle no 23 and no 21,&#160; 21 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Dissector 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MATERIALS FOR LUNG PROCUREMENT</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10 cc syringe</td>
			<td>BD DISCARDIT</td>
			<td>309110</td>
			<td></td>
		</tr>
		<tr>
			<td>Alprostadil 500 mcgs injectable solution</td>
			<td>PFIZER</td>
			<td></td>
			<td>Prostaglandin injected with lung preservation solution</td>
		</tr>
		<tr>
			<td>Disposable GIA cartridge Steril 6/Ca</td>
			<td>MEDTRONIC</td>
			<td>1141634</td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable GIA stapler 60/3.8 3/Ca</td>
			<td>MEDTRONIC</td>
			<td>2802122</td>
			<td>Stapler for trachea and bronquial division</td>
		</tr>
		<tr>
			<td>Foley catheter 18 Ch Folysil</td>
			<td>Folysil, Coloplast</td>
			<td>AA6118</td>
			<td>urinary catheter employed to canulated pulmonary veins for retrograde perfusion</td>
		</tr>
		<tr>
			<td>Lung preservation solution Perfadex 1000 mL</td>
			<td>Medisan, Uppsala, Sweeden</td>
			<td>19811 ( box of 10 units)</td>
			<td>Lung preservation solution</td>
		</tr>
		<tr>
			<td>Mersilk no 1, LR-60 CONV , 75 cm</td>
			<td>ETHICON</td>
			<td>W562H</td>
			<td>Silk curved suture for pericardium sutures</td>
		</tr>
		<tr>
			<td>Paediatric Venous cannula</td>
			<td>SORIN GROUP</td>
			<td>V132-12</td>
			<td>Cannula used for pulmonary artery cannulation</td>
		</tr>
		<tr>
			<td>Prolene 4/0</td>
			<td>ETHICON</td>
			<td>W8355</td>
			<td>polypropylene suture for purse string in pulmonary artery</td>
		</tr>
		<tr>
			<td>Scalpel blade no 11</td>
			<td>INTRAVEN</td>
			<td>150011</td>
			<td></td>
		</tr>
		<tr>
			<td>Sofsilk braided silk no 1 strands</td>
			<td>COVIDIEN</td>
			<td>L-12</td>
			<td>Silk strand to fix arterial cannula with the tourniquet</td>
		</tr>
		<tr>
			<td>Sofsilk braided silk no 3 strands</td>
			<td>COVIDIEN</td>
			<td>L-115</td>
			<td>Silk strand for vessel ligation</td>
		</tr>
		<tr>
			<td>Sterile bags</td>
			<td></td>
			<td></td>
			<td>To keep and store lungs.</td>
		</tr>
		<tr>
			<td>Straigth connector 1,4&#34;/1,4&#34; with luer lock</td>
			<td>ANDOCOR</td>
			<td>04CS0032</td>
			<td>Piece to connect pulmonary artery arterial cannula with preservation line and the three way stop-cock for prostaglandin</td>
		</tr>
		<tr>
			<td>Three-way stopcock</td>
			<td>BD CONNECTA</td>
			<td>394501</td>
			<td>Three way stop-cock to connect farterial cannula with pressure line</td>
		</tr>
		<tr>
			<td>Uromatic set for irrigation double lead</td>
			<td>MEDISAVE</td>
			<td>TRC4007N</td>
			<td>Irrigation system for lung preservation solution</td>
		</tr>
		<tr>
			<td>Uromatic set for irrigation single lead</td>
			<td>MEDISAVE</td>
			<td>TRC4002</td>
			<td>Irrigation system for lung preservation solution</td>
		</tr>
		<tr>
			<td>SURGICAL TOOLS FOR LUNG PROCUREMENT</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Aortic cross- clamp</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Battery-powered surgical saw</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cooley vascular clamp</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting forceps 18 cm and 27,9 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Finochietto sternal retractor</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Metzembaum scissors 20 cm and 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito forceps curved 12,5 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular clamps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SURGICAL TOOLS FOR ABDOMINAL ORGAN PROCUREMENT</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Adson articulated retractors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Allis forceps 16 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Aortic cross-clamps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Boyd scissors 17 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo needle holder</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cooley Vascular clamps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Crile forceps curved 18 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Davis retractor 24.5 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DeBakey dissecting forceps 19.7 cm adn 24.1 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DeBakey vascular clamps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting forceps 18 cm and 27.9 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Duval forceps 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Farabeuf retractors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Kidney Trays 300 cc and 500 cc</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Kocher forceps straigth 18 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Langenbeck retractors 21 cm and 23 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mayo scissors straigth and curved , 17 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito forceps straigth and curved, 12.5 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holders 15 cm, 18 cm, 23 cm and 23 cm.</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Pean forceps 16 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Potts scissors 19cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rochester forceps curved 24 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rochester forceps straigth 24 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Russ dissection forceps 15 cm and 20 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel handles</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Senn-mueller retractor 16 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

lung rapid recovery procurement combined with abdominal normothermic regional perfusion in controlled donation after circulatory death

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years confirm that the outcomes after lung transplantation from cDCD are similar to those from brain death donors; however, the utilization of lungs from asystole donors remains low. Several reasons may be involved: different legal frameworks among countries and centers with different premortem interventions, inadequate lung donor care before procurement, or even poor experience with cDCD procedures and protocols.
Initially, the rapid recovery technique was commonly employed for the procurement of thoracic and abdominal organs in cDCD, but, in the last decade, abdominal normothermic regional perfusion (ANRP) with extracorporeal membrane oxygenation devices has become a useful method to restore blood flow to abdominal organs, allowing their quality improvement and their functional assessment prior to transplantation. This makes the donation procedure more complex and generates doubts about injury to the grafts due to dual temperature.
The aim of this article is to describe a protocol based on a single center experience with Maastricht III donors combining lung cooling rapid recovery in the thorax and abdominal normothermic regional perfusion. Tips and tricks focused on premortem interventions and lung procurement procedure techniques are explained. This may help to minimize the reluctance among professionals to use this combined technique and encourage other donor centers to use it, despite the increased complexity of the procedure.

We performed a descriptive analysis of 30 lung transplants performed at University Hospital Marqu&#233;s de Valdecilla with lungs obtained from cDCD donors in the last 2 years, 2020 and 2021. Donor and recipient demographic characteristics, technical data, postoperative outcomes, and short-term results are presented here. These results are presented as absolute numbers and percentages for categorical variables and as measures of central tendency and dispersion for continuous variables. The Kolmogorov-Smirnov test was use...

Watch this Scientific Journal Video about Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death at JoVE.com

Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death

The protocol combines a lung cooling rapid recovery technique with abdominal normothermic regional perfusion for abdominal graft procurement in controlled asystole donors, which is a safe and useful method to expand the donor pool.

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years ...

Multi-organ recovery for controlled donation after cardiac death donors is a challenge. This method increases the complexity of the whole procedure, but it offers better results for the recipients. This method minimizes the ischemic damage produced by hypotension and cardiac arrest after the withdrawal of life-sustaining therapy in donors.To begin, intravenously administer a bolus of Heparin to the donor. Then prepare a sterile instrumentation table with all the necessary equipment, the electrocautery, and the suction system. Prepare a surgical field on the selected groin with disinfectant solution and sterile drapes.Make an eight to 10 centimeter longitudinal incision with a number 23 blade, and control the bleeding with electrocautery and Ligaclips. Separate the wound edges with a retractor, and proceed with dissection to expose the femoral artery and vein. Embrace femoral vessels with a silk strand to control the bleeding.Select the appropriate cannula diameters according to the vessel sizes as described in the text manuscript. Cannulate the femoral vein, introducing first a metal wire as a guide, followed by progressive dilators to finally introduce the cannula. Proceed similarly with the femoral artery using a double lumen cannula.Cut off a 10 centimeter piece of the ECMO input line. Insert a straight connector with a lure lock with a three-way stopcock assembled at one end of the piece, and connect the other end to the arterial cannula. Purge the ECMO lines.Employ an irrigation paired with saline to fill the lines while connecting with cannulas. Connect the output ECMO line to the venous cannula and the input ECMO line to the straight connector with the three-way stopcock previously assembled to the arterial cannula. The three-way stopcock can be used for purging the system.Keep the ECMO lines clamped. Fix both cannulas to the groin with size one silk sutures to avoid displacement during transfer. Placer a monitored pressure line in the femoral artery cannula and the donor's left radial artery.Next, initiate the aortic occlusion balloon placement by taking the distance between the xiphoid process of the donor and the distal end of the arterial cannula as a reference to determine the length of the catheter to be inserted to reach the thoracic descending aorta. Set a reference mark in the balloon with a silk suture or a marker. Introduce a metal wire guide through the free lumen of the femoral artery cannula.Continue with the catheter guided by the metal wire, and introduce it until the referenced mark. Confirm the correct position of the occlusion balloon. Check the correct function of the occlusion balloon by filling it with saline for four to five seconds using a 50 cubic centimeter cone syringe.Confirming that the arterial pressure from the femoral cannula disappears, while the pressure from the left radial artery is maintained. To perform a median sternotomy, proceed with a median vertical skin incision from the suprasternal notch to the tip of the xiphoid process. Extend the incision to the pectoral fascia and sternal periosteum using electrocautery.Divide the interclavicular ligament and create a plane by finger dissection behind the sternum, both at the level of the suprasternal notch and the xiphoid process. Divide the sternum with an electric saw. Place a sternal retractor and open it carefully, releasing the pericardium from the posterior surface of the sternum.Control any bleeding point with electrocautery. At the same time, ventilate the donor with 100%oxygen and a positive and expiratory pressure of five centimeter water. For bronchoscopy, introduce a flexible bronchoscope through the endotracheal tube.Open both the pleural cavities by longitudinal incisions in the mediastinal pleura. If any errors are encountered while examining the supra-aortic vessels with the occlusion balloon, retract the left lung medially to expose and clamp the thoracic aorta as low as possible under direct vision. Examine the lungs by performing visual and palpatory assessments, inspecting for bullae, contusion, atelectasis, pneumonia, and occult tumors.Deliver one liter of four degree Celsius saline in both pleural cavities. Reduce the inspired fraction of oxygen to 50%Open the pericardium with an inverted T incision. Retract laterally the edges of the pericardium with 2-0 silk sutures fixed to the skin with mosquito forceps to expose the heart structures.Place a 4-0 polypropylene purse-string suture on the main pulmonary artery below the bifurcation. Perform an arteriotomy with a number 11 blade, and dilate with curved mosquito forceps. Cannulate the pulmonary artery with a right angled, straight cannula clamped at the end.Connect the pulmonary artery cannula to the irrigation system line, assembling a straight connector with a lure lock and a three-way stopcock. Connect the irrigation system to the lung preservation solution and purge the lines. Start flushing 50 to 60 milliliters per kilogram of cold preservation solution in an antegrade fashion.Start flushing 500 micrograms of prostaglandin diluted in 100 milliliters of saline at the same time through the three-way stopcock. Open the left atrium directly to allow free drainage. If areas of atelectasis are found, recruit them with short inspiratory holds at 25 to 30 centimeter water pressure.Once preservation is finished, remove the pulmonary artery cannula, and announce to the rest of the team the intention to clamp the cava vein and start heart excision. Place a cross clamp in the inferior cava vein, making sure there is enough stump for the liver. Ligate and divide the inferior cava vein with a number three silk strand.Tie and divide the femoral superior cava vein from caudal to azygous with a number three silk strand. Secure the distal stump with a clamp. Leave the clamps remaining in the surgical field, being careful not to remove them by accident, as otherwise, the ECMO device and ARNP will be compromised.Excise the rest of the heart in a standard function. After heart excision, remove the lungs following the same procedure as with brain death donors. Divide the inferior pulmonary ligaments, open the posterior pericardium, and expose the esophagus.Free the posterior mediastinal attachments of the lung with blunt dissection, ensuring cautious hemostasis. Dissect the pulmonary arteries away from the aorta. Isolate the trachea above the carina and pass a TA stapler around.Inflate the lungs to 50 to 60%of the tidal volume before withdrawing the endotracheal tube and dividing the trachea. Remove any remaining attachments and extract the lung block from the donor. Carefully check the thoracic cavity to detect any bleeding points, since continuous blood loss may decrease the pump flow.The percentage of patients with interstitial lung disease was the highest, followed by chronic obstructive pulmonary disease, and bronchiectasis. Most recipients had a smoking history. Only 16.7%had systemic hypertension and 10%had diabetes mellitus.Pulmonary hypertension was present in 14 recipients. One recipient needed intraoperative extracorporeal life support during surgery with ECMO. The median cold ischemic time was 292.5 minutes for the first graft and 405 minutes for the second.There were no intraoperative deaths. Two recipients needed postoperative ECMO support due to primary graft dysfunction. The median time for postoperative intubation was 24 hours.ICU stay was 3.1 days and hospital stay was 18.9 days. Acute cellular rejection in the first three weeks was present in 12 recipients. There was no hospital mortality and 30 day survival was 100%The key points in this method are pre-mortem interventions, cannulation, aortic occlusion balloon placement, and the measures to avoid volume loss and pump flow during lung preservation and procurement.This method has been applied to pediatric donors with an increased rate of grafts utilization and excellent recipients survival.

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Ex Situ Normothermic Machine Perfusion of Donor Livers

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous perfusion of organs provides the opportunity to improve organ quality and allows ex situ viability assessment of donor livers prior to transplantation. This video article provides a step by step protocol for ex situ normothermic machine perfusion (37 &#176;C) of human donor livers using a device that provides a pressure and temperature controlled pulsatile perfusion of the hepatic artery and continuous perfusion of the portal vein. The perfusion fluid is oxygenated by two hollow fiber membrane oxygenators and the temperature can be regulated between 10 &#176;C and 37 &#176;C. During perfusion, the metabolic activity of the liver as well as the degree of injury can be assessed by biochemical analysis of samples taken from the perfusion fluid. Machine perfusion is a very promising tool to increase the number of livers that are suitable for transplantation.

Ex Situ Normothermic Machine Perfusion of Donor Livers

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion

There is currently a severe shortage of liver grafts available for transplantation. Novel organ preservation techniques are needed to expand the pool of donor livers. Machine perfusion of donor liver grafts is an alternative to traditional cold storage of livers and holds much promise as a modality to expand the donor organ pool. We have recently described the potential benefit of subnormothermic machine perfusion of human livers. Machine perfused livers showed improving function and restoration of tissue ATP levels. Additionally, machine perfusion of liver grafts at subnormothermic temperatures allows for objective assessment of the functionality and suitability of a liver for transplantation. In these ways a great many livers that were previously discarded due to their suboptimal quality can be rescued via the restorative effects of machine perfusion and utilized for transplantation. Here we describe this technique of subnormothermic machine perfusion in detail. Human liver grafts allocated for research are perfused via the hepatic artery and portal vein with an acellular oxygenated perfusate at 21 &#176;C.

We describe a method of ex vivo machine perfusion of human liver grafts at subnormothermic temperature (21 &#176;C).

There is currently a severe shortage of liver grafts available for transplantation.  Novel organ preservation techniques are needed to expand the pool of ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Pre-clinical Model of Cardiac Donation after Circulatory Death

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after circulatory death (DCD) is a solution to address this limited availability, but due to a period of prolonged warm ischemia and the risk of tissue injury, its routine use in cardiac transplantation is seldom seen. In this manuscript we provide a detailed protocol closely mimicking current clinical practices in the context of DCD with continuous monitoring of heart function, allowing for the evaluation of novel cardioprotective strategies and interventions to decrease ischemia-reperfusion injury.
In this model, the DCD protocol is initiated in anesthetized Lewis rats by stopping ventilation to induce circulatory death. When systolic blood pressure drops below 30 mmHg, the warm ischemic time is initiated. After a pre-set warm ischemic period, hearts are flushed with a normothermic cardioplegic solution, procured, and mounted onto a Langendorff ex vivo heart perfusion system. Following 10 min of initial reperfusion and stabilization, cardiac reconditioning is continuously evaluated for 60 min using intraventricular pressure monitoring. A heart injury is assessed by measuring cardiac troponin T and the infarct size is quantified by histological staining. The warm ischemic time can be modulated and tailored to develop the desired amount of structural and functional damage. This simple protocol allows for the evaluation of different cardioprotective conditioning strategies introduced at the moment of cardioplegia, initial reperfusion and/or during ex vivo perfusion. Findings obtained from this protocol can be reproduced in large models, facilitating clinical translation.

This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death.

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

Isolated Lung Perfusion System in the Rabbit Model

The isolated lung perfusion system has been widely used in pulmonary research, contributing to elucidate the lungs&#39; inner workings, both micro and macroscopically. This technique is useful in the characterization of pulmonary physiology and pathology by measuring metabolic activities and respiratory functions, including interactions between circulatory substances and the effects of inhaled or perfused substances, as in drug testing. While in vitro methods involve the slicing and culturing of tissues, the isolated ex vivo lung perfusion system allows to work with a complete functional organ making possible the study of a continuous physiological function while recreating ventilation and perfusion. However, it should be noted that the effects of the absence of central innervation and lymphatic drainage still have to be fully assessed. This protocol aims to describe the assembly of the isolated lung apparatus, followed by the surgical extraction and cannulation of lungs and heart from experimental lab animals, as well as to display the perfusion technique and signal processing of data. The average viability of the isolated lung ranges between 5-8 h; during this period, the pulmonary capillary permeability increases, causing edema and lung injury. The functionality of preserved pulmonary tissue is measured by the capillary filtration coefficient (Kfc), used to determine the extent of pulmonary edema through time.

The isolated rabbit lung preparation is a gold standard tool in pulmonary research. This publication aims to describe the technique as developed for the study of physiological and pathological mechanisms involved in airway reactivity, lung preservation, and preclinical research in lung transplantation and pulmonary edema.

The isolated lung perfusion system has been widely used in pulmonary research, contributing to elucidate the lungs&#39; inner workings, both micro and ...

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ quality exacerbated by excessive geographic transportation distance and stringent donor organ acceptance criteria pose limitations to current LTx efforts. Ex situ lung perfusion (ESLP) is an innovative technology that has shown promise in attenuating these limitations. The physiologic ventilation and perfusion of the lungs outside of the inflammatory milieu of the donor body affords ESLP several advantages over traditional cold static preservation (CSP). There is evidence that negative pressure ventilation (NPV) ESLP is superior to positive pressure ventilation (PPV) ESLP, with PPV inducing more significant ventilator-induced lung injury, pro-inflammatory cytokine production, pulmonary edema, and bullae formation. The NPV advantage is perhaps due to the homogenous distribution of intrathoracic pressure across the entire lung surface. The clinical safety and feasibility of a custom NPV-ESLP device have been demonstrated in a recent clinical trial involving extender criteria donor (ECD) human lungs. Herein, the use of this custom device is described in a juvenile porcine model of normothermic NPV-ESLP over a 12 h duration, paying particular attention to management techniques. Pre-surgical preparation, including ESLP software initialization, priming, and de-airing of the ESLP circuit, and the addition of anti-thrombotic, anti-microbial, and anti-inflammatory agents, is specified. The intraoperative techniques of central line insertion, lung biopsy, exsanguination, blood collection, cardiectomy, and pneumonectomy are described. Furthermore, particular focus is paid to anesthetic considerations, with anesthesia induction, maintenance, and dynamic modifications outlined. The protocol also specifies the custom device's initialization, maintenance, and termination of perfusion and ventilation. Dynamic organ management techniques, including alterations in ventilation and metabolic parameters to optimize organ function, are thoroughly described. Finally, the physiological and metabolic assessment of lung function is characterized and depicted in the representative results.

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

This paper describes a porcine model of negative pressure ventilation ex situ lung perfusion, including procurement, attachment, and management on the custom-made platform. Focus is made on anesthetic and surgical techniques, as well as troubleshooting.

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

Large-Animal Model of Donation after Circulatory Death and Normothermic Regional Perfusion for Cardiac Assessment

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ donor pool. However, the DCD process is associated with the risk of cardiac tissue injury due to the inevitable period of warm ischemia. Normothermic regional perfusion (NRP) allows for an in situ organ assessment, allowing the procurement of hearts determined to be viable. Here, we describe a clinically relevant large-animal model of DCD followed by NRP. Circulatory death is established in anesthetized pigs by stopping mechanical ventilation. After a preset warm ischemia period, an extracorporeal membrane oxygenator (ECMO) is used for a NRP period lasting at least 30 min. During this reperfusion period, the model allows the collection of various myocardial biopsies and blood samples for initial cardiac evaluation. Once NRP is weaned, biochemical, hemodynamic, and echocardiographic assessments of cardiac function and metabolism can be performed before organ procurement. This protocol closely simulates the clinical scenario previously described for DCD and NRP in heart transplantation and has the potential to facilitate studies aimed at decreasing ischemia-reperfusion injury and enhance cardiac functional preservation and recovery.

The protocol describes a large-animal (porcine) model of donation after circulatory death, followed by thoracoabdominal normothermic regional perfusion that closely simulates the clinical scenario in heart transplantation, and has the potential to facilitate therapeutic studies and strategies.

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ ...

Extracorporeal Cardiopulmonary Resuscitation in Animal Model After Cardiac Arrest

Pediatric Animal Model of Extracorporeal Cardiopulmonary Resuscitation After Prolonged Circulatory Arrest

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted worldwide per year. Comprehensive investigation of this disease requires appropriate and validated animal models. Piglets are commonly used for translational research due to their analogous anatomy and physiology. This work aimed to describe and validate a neonatal piglet model of cardiopulmonary bypass (CPB) with circulatory and cardiac arrest (CA) as a tool for studying severe brain damage and other complications of cardiac surgery. In addition to including a list of materials, this work provides a roadmap for other investigators to plan and execute this protocol. After experienced practitioners performed several trials, the representative results of the model demonstrated a 92% success rate, with failures attributed to small piglet size and variant vessel anatomy. Furthermore, the model allowed practitioners to select from a wide variety of experimental conditions, including varying times in CA, temperature alterations, and pharmacologic interventions. In summary, this method uses materials readily available in most hospital settings, is reliable and reproducible, and can be widely employed to enhance translational research in children undergoing heart surgery.

Author Spotlight: Simulating Pediatric Cardiac Surgery Using a Neonatal Piglet Model

This protocol describes a neonatal porcine model of cardiopulmonary bypass (CPB), with circulatory and cardiac arrest as a tool for studying severe brain damage and other complications secondary to CPB.

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted worldwide per year. Comprehensive ...

Study of Experimental Organ Donation Models for Lung Transplantation

Experimental models are important tools for understanding the etiological phenomena involved in various pathophysiological events. In this context, different animal models are used to study the elements triggering the pathophysiology of primary graft dysfunction after transplantation to evaluate potential treatments. Currently, we can divide experimental donation models into two large groups: donation after brain death and donation after circulatory arrest. In addition, the deleterious effects associated with hemorrhagic shock should be considered when considering animal models of organ donation. Here, we describe the establishment of three different lung donation models (post-brain death donation, post-circulatory death donation, and post-hemorrhagic shock donation) and compare the inflammatory processes and pathological disorders associated with these events. The objective is to provide the scientific community with reliable animal models of lung donation for studying the associated pathological mechanisms and searching for new therapeutic targets to optimize the number of viable grafts for transplantation.

The present study shows the establishment of three different lung donation models (post-brain death donation, post-circulatory death donation, and post-hemorrhagic shock donation). It compares the inflammatory processes and pathological disorders associated with these events.

Experimental models are important tools for understanding the etiological phenomena involved in various pathophysiological events. In this context, ...