Establishment, characterization and cryopreservation of Fars native goat fetal fibroblast cell lines

Abstract

Abstract Objective To biologically develop and evaluate the caprine fetal fibroblast cell cultures before and after freezing. Methods Goat fetuses (ages 51, 53 and 55 d) were collected from slaughterhouse. Their skin was cut into small pieces (1 mm 3 ) and cultured in DMEM and FBS. When reaching 80%–90% confluence, cells were passaged. Cells of the 8th passage were cultured in 24-well plates (1.5 × 10 5 cells/well) for 9 d and three wells were counted every day. The average cell counts at each time point were plotted against day number and the population doubling time (PDT) was determined. Then, 42 vials of cells (2 × 10 6 cells/mL) were frozen. Samples were thawed and cultured after 1 month. Cell viability and PDT were evaluated after thawing. Results After eight passages, the goat fetal fibroblast cells had a latent phase of about 48 h and after an exponential phase, cells entered the plateau phase on day 5. Before freezing, PDT was about 22 h and after thawing it was about 28 h. Conclusions The goat fetal fibroblast cell culture can be established using the adherent culture method and can be cryopreserved, too. After thawing, growth and viability indices of these cells were acceptable.