abberior dyes & labels
2024
Cells
In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue
Authors:
Nicole Procel, Karen Camacho, Elisabeth Verboven, Isabel Baroja, Priscila A. Guerrero, Hanne Hillen, Carlos Estrella-García, Nicole Vizcaíno-Rodríguez, Leticia Sansores-Garcia, Ana Santamaría-Naranjo, Andrés Romero-Carvajal, Andrés Caicedo, Georg Halder, Iván M. Moya
Keywords:
in vivo cell death tracking; Trypan Blue labeling; organ regeneration; anticancer therapy assessment; ischemia-reperfusion injury; cholangiocarcinoma; necrosis; apoptosis
Abstract:
Tracking cell death in vivo can enable a better understanding of the biological mechanisms underlying tissue homeostasis and disease. Unfortunately, existing cell death labeling methods lack compatibility with in vivo applications or suffer from low sensitivity, poor tissue penetration, and limited temporal resolution. Here, we fluorescently labeled dead cells in vivo with Trypan Blue (TBlue) to detect single scattered dead cells or to generate whole-mount three-dimensional maps of large areas of necrotic tissue during organ regeneration. TBlue effectively marked different types of cell death, including necrosis induced by CCl4 intoxication in the liver, necrosis caused by ischemia-reperfusion in the skin, and apoptosis triggered by BAX overexpression in hepatocytes. Moreover, due to its short circulating lifespan in blood, TBlue labeling allowed in vivo “pulse and chase” tracking of two temporally spaced populations of dying hepatocytes in regenerating mouse livers. Additionally, upon treatment with cisplatin, TBlue labeled dead cancer cells in livers with cholangiocarcinoma and dead thymocytes due to chemotherapy-induced toxicity, showcasing its utility in assessing anticancer therapies in preclinical models. Thus, TBlue is a sensitive and selective cell death marker for in vivo applications, facilitating the understanding of the fundamental role of cell death in normal biological processes and its implications in disease.