Improved BALB/c mice granulosa cell functions using purified alginate scaffold

Alginate, a non-toxic polysaccharide isolated from brown algae, is a widely used 3-dimensional (3D) porous scaffold for the granulosa cell and follicle encapsulation. However, impurities in commercial alginate can lead to alginate biocompatibility reduction. The aim of this study was to evaluate in vitro behavior of the granulosa cells seeded on the purified alginate in varying concentrations compared with matched non-purified ones. We produced a purified alginate using a simple and efficient method. Then, the granulosa cells from mice were isolated and seeded in various concentrations of (0.5%, 1% weight/volume) purified and non-purified alginate. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used on the 3rd, 5th and the 8th days of culture as an index of cell viability and proliferation. Furthermore, the secreted estradiol, progesterone and alkaline phosphatase enzyme (ALP) were measured in the granulosa cells culture media using radioimmunoassay kits. The cells cultured on purified and low concentration alginate showed a higher proliferation rate, sex hormone production and ALP activity. The results confirmed the impact of the alginate hydrogel properties on proliferative rate and function of granulosa cells in a 3D culture system.