Screening for Chemical Toxicity Using Cryopreserved Precision Cut Lung Slices
| Autor: | Christa Watson, Joseph D. Brain, Thomas C. Donaghey, Sylvia Rodrigues, Ramon M. Molina, Jamie H. Rosenblum Lichtenstein, Sumati Ram-Mohan, Ramaswamy Krishnan, Priscila de Moura Queiroz, Flavia Damiani |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Male Pathology medicine.medical_specialty Toxicity Assessment in Cryopreserved Lung Sections Cell Survival High-throughput screening Primary Cell Culture Lung injury Biology In Vitro Techniques Toxicology medicine.disease_cause Cryopreservation Monocytes Andrology Extracellular matrix 03 medical and health sciences chemistry.chemical_compound Chlorides Bone plate Toxicity Tests medicine Animals Humans Rats Wistar Lung Cells Cultured Membrane Potential Mitochondrial Toxin Epithelial Cells Glutathione Mice Inbred C57BL Oxidative Stress 030104 developmental biology medicine.anatomical_structure chemistry Organ Specificity Zinc Compounds |
| Zdroj: | Toxicological sciences : an official journal of the Society of Toxicology. 150(1) |
| ISSN: | 1096-0929 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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