An in vitro study of Hoechst 33342 redistribution and its effects on cell viability
Autor: | Grega Repovs, Mara Bresjanac, M Gorensek, Nina Mohorko, N Kregar-Velikonja |
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Rok vydání: | 2005 |
Předmět: |
0301 basic medicine
Bisbenzimide Programmed cell death Pathology medicine.medical_specialty Stromal cell Time Factors Cell Survival Health Toxicology and Mutagenesis Population Bone Marrow Cells Biology Toxicology Andrology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine medicine Animals Viability assay Rats Wistar education Incubation Cells Cultured Bone Marrow Transplantation Fluorescent Dyes education.field_of_study 030102 biochemistry & molecular biology Staining and Labeling General Medicine Rats Transplantation chemistry ROC Curve Apoptosis 030220 oncology & carcinogenesis Area Under Curve Benzimidazoles |
Zdroj: | Humanexperimental toxicology. 24(11) |
ISSN: | 0960-3271 |
Popis: | Although Hoechst 33342 (H342) is frequently used to label donor cells in cell transplantation research, it has been noted that it might secondarily label the host cells. Furthermore, its potential toxicity leading to cell death has been described. We studied the time course of H342 redistribution from the primary labeled rat bone marrow stromal cells (rBMSC) into the non-labeled rBMSC population over 7 days in culture; we evaluated the nuclear H342 fluorescence intensity as a possible criterion for distinguishing the primary from the secondary labeled cells, and determined the viability of rBMSC after an overnight incubation in 1 mg/mL of H342. H342 labeled / 50% of the initially non-labeled cells within the first 6 hours and almost 90% within a week.Nuclear fluorescence intensity was a reliable criterion for distinguishing primary and secondary labeled cells within the first 24 hours, but less so at later time points. The percentage of either apoptotic or necrotic cells did not rise acutely after the overnight incubation in 1 mg/mL of H342. Although a 12-hour incubation of rBMSC in 1 mg/mL of H342 did not cause acute cell death, H342 rapidly and extensively redistributed into non-labeled cells, which makes H342 a relatively unsuitable marker for cell transplantation research. |
Databáze: | OpenAIRE |
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