Despite their advantages, in vitro methods have certain limitations:
- Lack of Whole-Organism Complexity: In vitro systems may not fully replicate the complex interactions present in a living organism, leading to potential discrepancies in results.
- Limited Metabolic Capability: Some in vitro systems may not accurately mimic the metabolic processes of an entire organism, affecting the assessment of the substance's metabolic fate and potential toxicity.
- Regulatory Acceptance: While gaining acceptance, some regulatory bodies still require in vivo data for certain evaluations, limiting the sole reliance on in vitro methods.
