PMID: 11334972May 4, 2001Paper

Monitoring of cell viability and cell growth in a hollow-fiber bioreactor by use of the dye Alamar Blue

Journal of Immunological Methods
H GloecknerH D Lemke

Abstract

We describe a method for monitoring cell proliferation in a small-scale hollow-fiber bioreactor (culture volume: 1 ml) by use of the Alamar Blue dye. Alamar Blue is a non-fluorescent compound, which yields a fluorescent product after reduction, e.g. by living cells. In contrast to the MTT-assay, the Alamar Blue assay does not lead to cell death. However, when not removed from the cells, the Alamar Blue dye shows a reversible, time- and concentration-dependent growth inhibition as observed for the leukemic cell lines CCRF-CEM, HL-60 and REH. When applied in the medium compartment of a hollow-fiber bioreactor system, the dye is delivered to the cells across the hollow-fiber membrane, reduced by the cells and released from the cell into the medium compartment back again. Thus, fluorescence intensity can be measured in medium samples reflecting growth of the cells in the cell compartment. This procedure offers several advantages. First, exposure of the cells to the dye can be reduced compared to conventional culture in plates. Second, handling steps are minimized since no sample of the cells needs to be taken for readout. Moreover, for the exchange of medium, a centrifugation step can be avoided and the cells can be cultivated furt...Continue Reading

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Citations

Nov 5, 2005·Analytical and Bioanalytical Chemistry·Y DudalM Debroux
Oct 13, 2009·Biotechnology Letters·Marianne J EllisJulian B Chaudhuri
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