Meeting Abstract
Tissue culture is an important tool in biochemistry, cell biology, and cell physiology research. There are currently very few reported cell culture methods in marine invertebrates and even fewer tissue culture lines. We focused on using the model chordate Ciona intestinalis to develop tissue culture for the ascidian Ciona intestinalis. We used the Microtissues Inc. “3D Petri Dish” system, which consists of cells seeded into agarose microwells, to culture both ovarian and embryonic tissue from C. intestinalis. Comparison of the 3D culture and traditional cell culture on treated plastic showed that proliferation was greater and more sustained in the 3D system. This may be due to the 3D system allowing more natural cell-cell interactions. Culture medium was optimized using cell counts over time using different combinations of filtered sea water, deionized water, antibiotic, and growth additives such as fetal bovine serum and Ciona hemolymph. It was found that Ciona hemolymph was the best growth additive for optimal cell proliferation and is thought to also aid in the protection of the cell culture from protist contaminants that can take over the tissue culture. While the 2D cultures proliferated very slowly, if at all, and were overcome by contaminants, the 3D cultures are still healthy after 6 weeks.
