Establecimiento de cultivos primarios de células endometriales y oviductales bovinas

Abstract

Currently, primary cell cultures are widely used for various research purposes, thereby reducing the reliance on laboratory animals. The aim of this investigation is to develop a standard protocol for primary cell culture of bovine endometrial and oviductal cells suitable for the conditions of the School of Agricultural Science labs at Universidad de Cuenca. We utilized endometrial and oviductal bovine cells, subjecting them to different treatments including modifications to their culture medium such as bovine fetal serum (BFS), pyruvate, and antibiotics. A total of 5 uteri in the luteal phase were collected, from which cells were extracted for endometrial cultures (n=12) and oviductal cultures (n=12). Endometrial cells were cultured for 14 days, while oviductal cells were cultured for 8 days. Analyses including assessment of confluence, cellular vitality, and well contamination were conducted on specified days using vital staining and Gram staining. However, observation and culture medium changes were performed every 48 hours. The results indicated the influence of the applied treatments on cellular vitality for both endometrial and oviductal cells (p < 0.05). While cellular confluence and well contamination in endometrial cell cultures were not affected by the treatments applied (p > 0.05), confluence and well contamination in oviductal cells exhibited statistical differences among treatments (p < 0.05). In summary, an initial assay of a cell culture protocol was successfully conducted under the conditions of the Agricultural School laboratory. However, further refinement of this technique is necessary to enable the pursuit of more complex research inquiries using primary cell culture methods