Quantification of cyclobutane pyrimidine dimers in human epidermal stem cells.

M Ruetze, S Gallinat, H Wenck, A Knott

R&D, Skin Research Center, Beiersdorf AG, Hamburg, Germany.

Methods in molecular biology (Clifton, N.J.) 2013

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The common procedures that are used to quantify cyclobutane pyrimidine dimers (CPD) comprise the extraction of cellular DNA followed by the detection of this nucleic acid modification by immunoblotting or electrophoretic methods. Consequently, these approaches provide an averaged damage intensity value of a whole population of cells and are not applicable to studies where a small subgroup such as somatic stem cells are intended to be investigated and the individual cellular damage is of interest. Here, we describe a strategy to isolate epidermal stem cells from minimum human epidermis samples and a subsequent immunocytochemical quantification of cellular CPDs. Besides the determination of the DNA damage status, this technique allows for the examination of cellular CPD intensity distributions.

Citation

M Ruetze, S Gallinat, H Wenck, A Knott. Quantification of cyclobutane pyrimidine dimers in human epidermal stem cells. Methods in molecular biology (Clifton, N.J.). 2013;989:99-108