| Accuracy of three techniques to determine cell viability in 3D tissues or scaffolds. | |
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MedLine Citation:
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PMID: 18800876 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Several different assays are commonly used to evaluate survival of cells inside tissues or three-dimensional carriers, but their accuracy and reliability have not been evaluated. Here, we compare three methods for cell viability (CV) determination: (i) lactate dehydrogenase (LDH) staining on cryosections, (ii) calcein AM/ethidium homodimer-1 (CaAM/EthH) staining, and (iii) carrier digestion and trypan blue (TB) assay. Living and dead cell populations were generated from bovine chondrocytes and combined to produce approximately 0%, 25%, 50%, 75%, and 100% CV mixtures. CV ratios were measured with TB assay (MIX) before seeding cells into fibrin carriers. CV was then determined using the three methods (n = 5/method). Custom-written macros were used to process LDH- and CaAM/EthH-stained images, and hand counting with hemocytometer was used for the TB method. Absolute error and intraclass correlation (ICC) were used for accuracy and reliability evaluation. All methods estimated CV values close to MIX values. TB method was the most accurate (ICC = 0.99) followed by CaAM/EthH (ICC = 0.98) and LDH (ICC = 0.97). As for absolute quantification of living and dead cells, TB and LDH methods performed well (ICC = 0.75-0.96), whereas CaAM/EthH largely overestimated cell numbers (living, ICC = 0.30; dead, ICC = 0.30). Although TB was the most accurate, LDH and CaAM/EthH provide valuable information on cell shape and spatial distribution of cells in tissue or a scaffold. |
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Authors:
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Benjamin Gantenbein-Ritter; Esther Potier; Stephan Zeiter; Marije van der Werf; Christoph M Sprecher; Keita Ito |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Tissue engineering. Part C, Methods Volume: 14 ISSN: 1937-3384 ISO Abbreviation: - Publication Date: 2008 Dec |
Date Detail:
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Created Date: 2008-12-05 Completed Date: 2009-05-11 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101466663 Medline TA: Tissue Eng Part C Methods Country: United States |
Other Details:
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Languages: eng Pagination: 353-8 Citation Subset: IM |
Affiliation:
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AO Research Institute, AO Foundation, Davos, Switzerland. benjamin.gantenbein@aofoundation.org |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cattle Cell Count / methods Cell Culture Techniques / methods* Cell Survival* Cells, Cultured Ethidium / chemistry Fibrin / chemistry Fluoresceins / chemistry Imaging, Three-Dimensional L-Lactate Dehydrogenase / metabolism Reproducibility of Results Tissue Engineering / methods* Trypan Blue / chemistry Trypsin / chemistry |
| Chemical | |
Reg. No./Substance:
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0/Fluoresceins; 1461-15-0/fluorexon; 3546-21-2/Ethidium; 72-57-1/Trypan Blue; 9001-31-4/Fibrin; EC 1.1.1.27/L-Lactate Dehydrogenase; EC 3.4.21.4/Trypsin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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