Toxicology Studies in hPT Primary Cell Cultures

Responses of primary cultures of hPT cells to the nephrotoxic metabolite of TCE 5-(l,2-dichlorovinyl)-L-cysteine (DCVC) provide an illustration of this principle. We showed that primary cultures of hPT cells incubated with DCVC undergo necrotic cell death at relatively high concentrations ( 100 pM) and long incubation times ( 24h), whereas incubation with relatively low concentrations ( 100 pM) for relatively short periods of time ( 8h) can result in apoptosis (Lash et al., 2(X)1). An increase in the proportion of hPT cells in S-phase (as assessed by flow cytometry) and rates of DNA synthesis (as assessed by incorporation of [ H]-dTTP) also occurred with DCVC treatment and followed similar patterns as apoptosis. This suggests that such low-dose, early responses can only occur when the hPT cell population still has adequate energy and viability. 

I Interaction with cellular macromolecules Toxic insult.  

Numerous methods are available to identify and quantify the extent of mitochondrial dysfunction in hPT cells, such as measurements of ATP concentrations by colorimetric or luminescence-based assays, rates of substrate-dependent 

As described previously, a panel of biomarkers of effect, including Kim-1, CysC, and NGAL, has recently been approved by the US Food and Drug Administration for detection of adverse effects on kidney function (Hoffmann etal., 2010). Kim-1 is an excellent case in point and has been used in primary cultures of hPT cells to detect cytotoxicity (Huang et al 2015). Thus, these highly sensitive and renal-specific biomarkers can be monitored in a preclinical model that is highly relevant to humans. 

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