Abstract
Endothelialization of the blood contacting surfaces of blood-contacting medical devices, such as cardiovascular prostheses or biohybrid oxygenators, represents a plausible strategy for increasing their hemocompatibility. Nevertheless, isolation and expansion of autologous endothelial cells (ECs) usually requires multiple processing steps and time to obtain sufficient cell numbers. This excludes endothelialization from application in acute situations. Off-the-shelf availability of cell-seeded biohybrid devices could be potentially facilitated by hypothermic storage. In this study, the survival of cord-blood-derived endothelial colony forming cells (ECFCs) that were seeded onto polymethylpentene (PMP) gas-exchange membranes and stored for up to 2 weeks in different commercially available and commonly used preservation media was measured. While storage at 4°C in normal growth medium (EGM-2) for 3 days resulted in massive disruption of the ECFC monolayer and a significant decline in viability, ECFC monolayers preserved in Chillprotec could recover after up to 14 days with negligible effects on their integrity and viability. ECFC monolayers preserved in Celsior, HTS-FRS, or Rokepie medium showed a significant decrease in viability after 7 days or longer periods. These results demonstrated the feasibility of hypothermic preservation of ECFC monolayers on gas-exchange membranes for up to 2 weeks, with potential application on the preservation of pre-endothelialized oxygenators and further biohybrid cardiovascular devices.
Highlights
Thrombus formation due to the incomplete hemocompatibility of blood contacting surfaces represents a high risk factor in the long-term application of medical devices, such as vascular prostheses or extracorporeal membrane oxygenators (ECMO)
While there have been a number of studies reporting on the benefit of surface endothelialization of various blood contacting medical devices, including vascular prostheses,[1] circulatory assist devices,[2] stents, and gas-exchange membranes of ECMO devices,[3,4] the clinical applicability of such biohybrid devices, especially in acute situations, has not been extensively addressed
endothelial colony forming cells (ECFCs) have been shown to possess a high proliferation capacity, enabling the generation of large cell numbers that would be necessary for the seeding of large areas, as in the case of the biohybrid lung (>2.5 m2 surface area)
Summary
Thrombus formation due to the incomplete hemocompatibility of blood contacting surfaces represents a high risk factor in the long-term application of medical devices, such as vascular prostheses or extracorporeal membrane oxygenators (ECMO). Recent work has demonstrated the successful generation of MHC-silenced ECFCs by knocking down the β-2-microglobulin subunit of the MHC class-I complex, which has been reported to cloak these cells from the host’s immune system.[5] immunophenotype-matched allogeneic ECs and iPSC-derived-ECs could be considered for biohybrid devices.[6] These cell sources could be potentially used in the biomanufacturing of biohybrid devices with off-the-shelf availability, without relying on an autologous cell source Along these lines, the aim of this study was to investigate the feasibility of preserving endothelialized polymethylpentene (PMP) gas-exchange membranes, currently used in ECMO devices, under hypothermic conditions (4°C) for up to 2 weeks in different hypothermic preservation media. A 2-week shelf-life for a biohybrid ECMO would represent an important first step toward the clinical adaptation of the technology, facilitating its logistics and allowing for the necessary safety and quality assessment.[7]
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