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Stem Cell analysis: Functional analysis
Cytosolic aldehyde dehydrogenase (ALDH) is responsible for oxidising intracellular aldehydes and is involved in various cellular metabolic pathways. High levels of this enzyme are found in stem cells and are thought to be in part responsible for their increased drug resistance. It is possible to use a simple flow cytometric technique to identify cells that have functional ALDH - this is achieved by adding a non-fluorescent substrate that is cleaved in the presence of the enzyme to form a fluorescent product. The kit we use for this is AldeFluor - no antibody staining is required, it allows dead cells to be excluded from analysis and can potentially be used with other surface markers.
The example below, courtesy of Daniel Pearce (Haematopoietic Stem Cell Lab), shows human cord blood - the bright ALDH staining exhibits a low/medium side scatter population that, when superimposed over the whole mononuclear cell population (grey), appear to have the scatter characteristics of stem/progenitor cells.
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