Even though the efficacy of photodynamic therapy (PDT) for treating premalignant and malignant lesions has been demonstrated, resistant tumor cells to this therapy occasionally appear. Here, we describe the published methods to isolate resistant cancer cells to PDT and propose new procedures that may be used, as laboratory models allow a better understanding of resistance mechanisms. For this purpose, the treatment conditions, the photosensitizer (PS) or pro-drug, the cell line and the final selection - clonal of total population - must be taken into account. In general, high and repeated treatment doses are used. The resistant cell population characterization may include cell morphology, response to PDT, expression of death proteins or survival related genes and cell proliferation analysis. In addition, in vivo models such as the resistant cell transplantation to mice, allow evaluating tumorigenicity and aggressiveness, leading to the determination of the in vivo resistance. Summarizing, in order to improve clinical results, cellular models can help understand PDT-resistance mechanisms in vivo and in vitro.