Department of Biology, Faculty of Science, Silpakorn University, Nakorn Pathom, Thailand
The study on effect of porcine follicular fluid (pFF) with difference concentration on in vitro cell viability was done using MTT assay and analyzed by one-way ANOVA. Granulosa cells (GC) from medium size follicle (4-6 mm in diameter) were cultured in M-199 supplement with 10% FCS and 1 μg/mL LH, 1 μg/mL for 48 h. After culture time, 70-80% GC morphology were elongated from round shape to fibroblast-like morphology. Granulosa cells were treated with pFF of large follicle group (= 7 mm in diameter) at 2, 4, 20, 40, 200, 400, 500, 600 μg protein concentration for 24 hours. Results reviewed that, at concentration 2 μg, cell viability start increased slightly. At 500 μg concentration show the highest cell viability which higher than positive control (medium supplement with 10% FCS) and significantly difference with control group (serum free medium) (P<0.05). At this concentration, GC were doubles elongated and expanded more than control group and positive control (50-60%). The further study is to use pFF for supplement instead of serum and hormone in culture medium. This finding was an initial tool for use pFF as substance to promote cells growth, cells viability and be useful in biotechnology research.
Keywords: Primary cell culture, granulosa cells, follicular fluid, MTT assay, porcine.