Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Jun 1;14:12618. doi: 10.1038/s41598-024-63041-8

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Perfusion system used for loading and recovery, and detailed supercooling protocol. (a) Perfusion system. (i) The VCA is placed on a rack inside a stainless-steel bowl containing the perfusate solution. (ii) A peristaltic pump generates a flow in the silicon tubing to reach (iv) the Hollow-fiber membrane oxygenator, which also traps potential air bubbles. (v) A pressure sensor is connected to the closed system and to (iii) a pressure monitor, allowing for continuous monitoring. (vi) The oxygenated perfusate reaches the VCA through the arterial cannula. (vii) A hot plate stirrer keeps the perfusate at the desired temperature. White arrow = thermometer. (b) Supercooling protocol. The VCA is first loaded with 3-OMG as an intracellular CPA using SNMP (21 °C), before incremental switching to the extracellular CPA cocktail while decreasing the temperature until reaching 4 °C. The limb is then placed for 48 h in a sterile bag full of the extracellular CPA cocktail at − 5 °C in a cooler. The recovery is performed using SNMP with normal Steen + solution with incremental unloading of the extracellular CPA during gradual rewarming. The SNMP is pursued 2 h before switching to whole autologous blood warmed at 37 °C to perform the 2 h NMP phase. 3-OMG 3-O-methyl-glucose, CPA cryoprotective agent, HTK histidine-tryptophan-ketoglutarate, NMP normothermic machine perfusion, PEG polyethylene glycol, SNMP subnormothermic machine perfusion.