TY - JOUR
T1 - Zebrafish as a high throughput model for organ preservation and transplantation research
AU - da Silveira Cavalcante, Luciana
AU - Lopera Higuita, Manuela
AU - González-Rosa, Juan Manuel
AU - Marques, Beatriz
AU - To, Samantha
AU - Pendexter, Casie A.
AU - Cronin, Stephanie E. J.
AU - Gopinathan, Kaustav
AU - de Vries, Reinier J.
AU - Ellett, Felix
AU - Uygun, Korkut
AU - Langenau, David M.
AU - Toner, Mehmet
AU - Tessier, Shannon N.
N1 - Funding Information: This research was funded through SNT from the US National Institutes of Health (NIH), including K99/R00 HL1431149 and R01HL157803, American Heart Association (AHA; 18CDA34110049), Harvard Medical School Eleanor and Miles Shore Fellowship, and the Claflin Distinguished Scholar Award on behalf of the Massachusetts General Hospital (MGH) Executive Committee on Research. We also thankfully acknowledge support for MT (National Science Foundation under Grant No. EEC 1941543), DML (NIH R24OD031955; MGH Research Scholars), and JMGR (NIH R01HL164749‐01; AHA 19CDA34660207; Hassenfeld Scholar Award; Corrigan‐Minehan Heart Center of MGH SPARK Award). Publisher Copyright: © 2023 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.
PY - 2023/10/1
Y1 - 2023/10/1
N2 - Despite decades of effort, the preservation of complex organs for transplantation remains a significant barrier that exacerbates the organ shortage crisis. Progress in organ preservation research is significantly hindered by suboptimal research tools that force investigators to sacrifice translatability over throughput. For instance, simple model systems, such as single cell monolayers or co-cultures, lack native tissue structure and functional assessment, while mammalian whole organs are complex systems with confounding variables not compatible with high-throughput experimentation. In response, diverse fields and industries have bridged this experimental gap through the development of rich and robust resources for the use of zebrafish as a model organism. Through this study, we aim to demonstrate the value zebrafish pose for the fields of solid organ preservation and transplantation, especially with respect to experimental transplantation efforts. A wide array of methods were customized and validated for preservation-specific experimentation utilizing zebrafish, including the development of assays at multiple developmental stages (larvae and adult), methods for loading and unloading preservation agents, and the development of viability scores to quantify functional outcomes. Using this platform, the largest and most comprehensive screen of cryoprotectant agents (CPAs) was performed to determine their toxicity and efficiency at preserving complex organ systems using a high subzero approach called partial freezing (i.e., storage in the frozen state at −10°C). As a result, adult zebrafish cardiac function was successfully preserved after 5 days of partial freezing storage. In combination, the methods and techniques developed have the potential to drive and accelerate research in the fields of solid organ preservation and transplantation.
AB - Despite decades of effort, the preservation of complex organs for transplantation remains a significant barrier that exacerbates the organ shortage crisis. Progress in organ preservation research is significantly hindered by suboptimal research tools that force investigators to sacrifice translatability over throughput. For instance, simple model systems, such as single cell monolayers or co-cultures, lack native tissue structure and functional assessment, while mammalian whole organs are complex systems with confounding variables not compatible with high-throughput experimentation. In response, diverse fields and industries have bridged this experimental gap through the development of rich and robust resources for the use of zebrafish as a model organism. Through this study, we aim to demonstrate the value zebrafish pose for the fields of solid organ preservation and transplantation, especially with respect to experimental transplantation efforts. A wide array of methods were customized and validated for preservation-specific experimentation utilizing zebrafish, including the development of assays at multiple developmental stages (larvae and adult), methods for loading and unloading preservation agents, and the development of viability scores to quantify functional outcomes. Using this platform, the largest and most comprehensive screen of cryoprotectant agents (CPAs) was performed to determine their toxicity and efficiency at preserving complex organ systems using a high subzero approach called partial freezing (i.e., storage in the frozen state at −10°C). As a result, adult zebrafish cardiac function was successfully preserved after 5 days of partial freezing storage. In combination, the methods and techniques developed have the potential to drive and accelerate research in the fields of solid organ preservation and transplantation.
KW - cardiac graft
KW - cryobiology
KW - partial freezing
KW - solid organ preservation
KW - zebrafish
UR - http://www.scopus.com/inward/record.url?scp=85171333590&partnerID=8YFLogxK
U2 - https://doi.org/10.1096/fj.202300076R
DO - https://doi.org/10.1096/fj.202300076R
M3 - Article
C2 - 37718489
SN - 0892-6638
VL - 37
JO - FASEB Journal
JF - FASEB Journal
IS - 10
M1 - e23187
ER -