title: Screening for Chemical Toxicity Using Cryopreserved Precision Cut Lung Slices.
abstract: To assess chemical toxicity, current high throughput screening (HTS) assays rely primarily on in vitro measurements using cultured cells. Responses frequently differ from in vivo results due to the lack of physical and humoral interactions provided by the extracellular matrix, cell-cell interactions, and other molecular components of the native organ. To more accurately reproduce organ complexity in HTS, we developed an organotypic assay using the cryopreserved precision cut lung slice (PCLS) from rats and mice. Compared to the never-frozen PCLS, their frozen-thawed counterpart slices showed viability or metabolic activity that is decreased to an extent comparable to that observed in other cryopreserved cells and tissues, but shows no differences in further changes in cell viability, mitochondrial integrity, and glutathione activity in response to the model toxin zinc chloride (ZnCl2). Notably, these measurements were successfully miniaturized so as to establish HTS capacity in a 96-well plate format. Finally, PCLS responses correlated with common markers of lung injury measured in lavage fluid from rats intratracheally instilled with ZnCl2. In summary, we establish that the cryopreserved PCLS is a feasible approach for HTS investigations in predictive toxicology.
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Christa Y
authors.initials: CY
authors.lastname: Watson
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Flavia
authors.initials: F
authors.lastname: Damiani
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and Department of Emergency Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02115.
authors.forename: Sumati
authors.initials: S
authors.lastname: Ram-Mohan
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Rodrigues
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Priscila
authors.initials: P
authors.lastname: de Moura Queiroz
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Donaghey
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Jamie H
authors.initials: JH
authors.lastname: Rosenblum Lichtenstein
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Brain
authors.affiliation: Department of Emergency Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02115.
authors.forename: Ramaswamy
authors.initials: R
authors.lastname: Krishnan
authors.affiliation: *Molecular and Integrative Physiological Sciences Program, Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115 and rmolina@hsph.harvard.edu.
authors.forename: Ramon M
authors.initials: RM
authors.lastname: Molina
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002712:Chlorides / Q000633:toxicity; D015925:Cryopreservation; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D009928:Organ Specificity; D018384:Oxidative Stress / Q000187:drug effects; D061251:Primary Cell Culture; D017208:Rats, Wistar; D018675:Toxicity Tests / Q000379:methods*; D017967:Zinc Compounds / Q000633:toxicity
chemical_list: D002712:Chlorides; D017967:Zinc Compounds; C016837:zinc chloride
keywords: cryopreservation; high throughput screening; organotypic; precision cut lung slices; toxicology; zinc chloride