Comparison of Two Renal Cell Lines (NRK-52E and LLC-PK1) as Late Stage Apoptosis Models

Alexandra H. Heussner, Daniel R. Dietrich

Human and Environmental Toxicology, University of Konstanz, Konstanz, Germany.

DOI: 10.4236/ojapo.2013.23004                

Abstract

Renal cell lines are frequently used models in toxicology. The aim of the experiments described here was to investigate the suitability of two of those renal cell lines, namely NRK-52E and LLC-PK1, as models for mid to late stage apoptosis under standard cell culture conditions; the latter means that testing was performed in the presence of serum in the culture media. Seven known inducers of apoptosis already positively tested by other investigators were chosen as test substances using chromatin condensation (Hoechst staining) as endpoint. These substances were cadmium chloride (CdCl2), dithiothreitol (DTT), sodium chloride (NaCl), mercuric chloride (HgCl2), tributyltin oxide (TBT-O), tributyltin chloride (TBT-Cl) and staurosporine. From these, only TBT-O, TBT-Cl and staurosporine induced morphological features typical of apoptosis in LLC-PK1 cells. Morphologically discerned apoptosis was confirmed by DNA fragmentation (DNA laddering assay) analysis. LLC-PK1 cells, but not NRK-52E cells, were shown to be suitable models of mid to late stage apoptosis under the conditions employed. TBT-O, TBT-Cl and staurosporine were shown to be suitable positive controls for apoptosis in renal cells in vitro.

Keywords

Renal Cell Lines; Hoechst 33342; DNA Laddering

Conflicts of Interest

The authors declare no conflicts of interest.

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