Quantitative assessment of marine sponge cells in vitro: Development of improved growth medium

As sources of natural products with potential human therapeutic value, mari ne sponges are important subjects for cell culture studies. A critical comp onent of any cell culture system is its growth medium. Proceeding from the hypotheses that the thawed, cryopreserved, primary cells would display dete ctable differential responses and that those responses could be comparative ly; quantified, this study has established that multiwell screening assays are useful tools for improving medium formulations in cell cultures of the marine sponge, Teichaxinella morchella. Fluorescent probe signals were corr elated with known cell densities and viabilities in a 96-well format. analy sis of variance and post-test methods were applied to judge the significanc e of signal differences seen in a variety of medium formulations. Results f rom a series of experiments suggested that reducing glutamine and selenium concentrations in the standard medium would result in greater DNA, protein, and esterase activity signals. This was confirmed by the direct comparison of the standard and improved medium formulations. Significantly higher pro tein content and esterase activity were associated with the improved medium . DNA content was also higher, though not significantly. The result is a ne w medium formulation that may be more able to support cell growth and divis ion, providing an improved cell culture system for marine sponge cell studi es. The assays can be used in additional studies to further improve the in vitro conditions for marine sponge cell culture.