Abstract Background and purpose: The most common serum used in culture media is fetal bovine serum (FBS). Given that FBS is costly, this study aimed at evaluating the hydatid cyst fluid (HCF) as a substitute for FBS in culture medium of fibroblast cells isolated from rat. Materials and methods: In this experimental study, skin dermal area from inguinal and abdomen of rat were used for fibroblast preparation. Fibroblasts cells were isolated using trypsin enzyme and then were cultured in RPMI medium (including 20% FBS). After sufficient cell growth, they were isolated and cultured in 24-well microtiter plate in three successive concentrations as follows: pure RPMI and RPMI medium supplemented with 10% and 20% FBS as control groups, and RPMI medium supplemented with 10% and 20% HCF. Then survival rate and proliferation level were checked in three successive days using invert microscope (morphology evaluation) and trypan blue 0.4% as vital dye (quantitative evaluation), respectively. Results: Proliferation and growth of fibroblast cells in RPMI medium supplemented with 10% and 20% FBS concentrations were normal during the three days with 95% survival rate. In contrast, fibroblast cells in RPMI medium supplemented with 10% and 20% HCF concentrations were converted from normal to spherical form over the three days and 80 % of them were alive. Conclusion: The results indicated that we can apply HCF as an alternative for FBS up to 72h in RPMI medium. It can also be used as transport or short maintenance media for fibroblast cells.

Keywords: Key words: Fibroblast cells, RPMI medium, hydatid cyst fluid (HCF), fetal bovine serum (FBS), rat

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