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Model-Guided Design and Optimization of CPA Perfusion Protocols for Whole Organ Cryopreservation.

ABSTRACT: Vitrification could enable long-term organ preservation, but only after loading high-concentration, potentially toxic cryoprotective agents (CPAs) by perfusion. In this paper, we combine a two-compartment Krogh cylinder model with a toxicity cost function to theoretically optimize the loading of CPA (VMP) in rat kidneys as a model system. First, based on kidney perfusion experiments, we systematically derived the parameters for a CPA transport loading model, including the following: V_b = 86.0% (r_a = 3.86 μm), L_p = 1.5 × 10^-14 m³/(N·s), ω = 7.0 × 10^-13 mol/(N·s), σ = 0.10. Next, we measured the toxicity cost function model parameters as α = 3.12 and β = 9.39 × 10^-6. Combining these models, we developed an improved kidney-loading protocol predicted to achieve vitrification while minimizing toxicity. The optimized protocol resulted in shorter exposure (25 min or 18.5% less) than the gold standard kidney-loading protocol for VMP, which had been developed based on decades of empirical practice. After testing both protocols on rat kidneys, we found comparable physical and biological outcomes. While we did not dramatically reduce toxicity, we did reduce the time. As our approach is now validated, it can be used on other organs lacking defined toxicity data to reduce CPA exposure time and provide a rapid path toward developing CPA perfusion protocols for other organs and CPAs.

SUBMITTER: Han Z

PROVIDER: S-EPMC10518287 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature

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Model-Guided Design and Optimization of CPA Perfusion Protocols for Whole Organ Cryopreservation.

Han Zonghu Z Rao Joseph Sushil JS Ramesh Srivasupradha S Hergesell Jan J Namsrai Bat-Erdene BE Etheridge Michael L ML Finger Erik B EB Bischof John C JC

Annals of biomedical engineering 20230623 10

Vitrification could enable long-term organ preservation, but only after loading high-concentration, potentially toxic cryoprotective agents (CPAs) by perfusion. In this paper, we combine a two-compartment Krogh cylinder model with a toxicity cost function to theoretically optimize the loading of CPA (VMP) in rat kidneys as a model system. First, based on kidney perfusion experiments, we systematically derived the parameters for a CPA transport loading model, including the following: V<sub>b</sub ...[more]

PMID: 37351756

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Project description:BackgroundThe aim of this study was to develop an adipose tissue (AT) cryopreservation protocol that is effective, simple, and maintains the functionality and viability of AT after thawing and transplantation.MethodsTwo cryopreservation temperatures (T°), -20°C and -80°C, and two cryoprotective agents (CPAs), trehalose and hydroxyethyl starch (HES), were compared first in an experimental study, using a slowfreezing protocol. The five experimental groups were the following: (a) Fresh AT (control group), (b) T = -20°C, 10%HES, (c) T = -80°C, 10%HES, (d) T = -20°C, 0.35M trehalose, (e) T = -80°C, 0.35M trehalose. We evaluated the morphology (histological studies) and tissue viability by glyceraldehyde 3-phosphate dehydrogenase (GAPDH) genic expression. Based on the results of the preliminary study, an in vivo study was performed, choosing as cryopreservation T° -20°C. HES and trehalose were compared as cryoprotective agents and with a control group (fresh AT). AT grafts were transplanted into immunodeficient mice. After 1 month of inoculation, animals were euthanized and samples were recovered. Samples were weighted and processed for histological study, viability study (GAPDH genic expression), and vascularization study (VEGF genic expression).ResultsThe initial histological study demonstrated that all AT cryopreserved group samples showed typical histological features of AT, similar to that of the control group. Statistically significant differences were not observed (P > 0.05) in GAPDH expression between different groups related to temperature or CPA. Referring to the in vivo studies, cryopreserved groups showed good take of the graft and normal AT architectural preservation, as well as a clear vascular network. Statistically significant differences were not found (P > 0.05) with regard to graft take (%), GAPDH, or VEGF expression.ConclusionSlow freezing at -20°C using trehalose, and -20°C using HES as cryoprotective agents are both straightforward and easy AT cryopreservation procedures, with results similar to those of fresh AT in terms of tissue viability and morphohistological characteristics.

Dataset Information

Model-Guided Design and Optimization of CPA Perfusion Protocols for Whole Organ Cryopreservation.

Publications

Model-Guided Design and Optimization of CPA Perfusion Protocols for Whole Organ Cryopreservation.

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