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Background: Lead (Pb) is one of the most difficult metals to investigate in laboratory experiments because it is very easily precipitates or forms complex ions. Many experiments were conducted with Pb and observed unique chemical properties of this metal. Objective: The present investigation was designed to evaluate the cytotoxicity of Pb exposer on pheochromocytoma (PC12) cells in different condition and media to admit the difficulties facing the researchers in bioavailability of Pb in media and prevent precipitation formation. Materials and Methods: Pb from 0.01 to 100 μM had been used on PC12 cells cultured and treated in different conditions for 6, 12, 24, and 48 h. Resazurin assay was used to assess the cell viability.Result: As numerous studies have demonstrated a possible mechanism for Pb-induced neurotoxicity using in vitro model, the current study is the first study which explained the complexity of Pb participations in media that facing the authors and what the reason for that and how we can solve this problem to make pb more bioavailable in the media. Conclusion: Use of phosphate buffer in experiments with Pb is not acceptable because all Pb can be precipitated. However, it could be possible to use the standard media, but for the time of treatment with Pb phosphates must be removed from the media.

Diagnosis of the problems which facing the researchers during studying In vitro lead toxicity