Primary Cultures of hPT Cells

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. 

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. 

TABLE 10.4 Panel of Assays that Can Be Conducted in Primary Cultures of hPT Cells for Drug Design and Screening and Investigation of Mechanism of Action... 

Potential limitations to the use of primary cultures of hPT cells include the need to consider the potential for genetic... 

In summary, primary cultures of hPT cells represent a unique and highly relevant experimental model for the study of drug metabolism, transport, mechanism of action, and nephrotoxicity. Although there exist the usual cautions with primary cell culture models, and cell culture models in general, once technique is mastered and appropriate conditions are used to minimize potential issues with the culture process, the primary hPT cells have numerous advantages over other models for obtaining information about drug action that is directly relevant to humans. 

Advantages and Limitations of Performing Studies in hPT Primary Cell Cultures... 

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