The Benefits of the Fish Gill Cell Culture System include:
This Fish Gill Cell Culture System cultures the primary gill cells of the freshwater rainbow trout on a flat permeable membrane. This procedure, known as the double seeded insert (DSI) technique, produces a heterogeneous gill epithelium that is the most realistic in vitro alternative to the gill organ.
The key features of FiGCS is that it is an electrically tight gill epithelium, which contains different cell types, including pavement cells and mitochondria rich cells, present in the intact gill, that as well as physiologically resembling the gill tissue, can tolerate freshwater akin to the normal fish gill. These characteristics are fundamental in recreating an in vitro freshwater gill epithelium that can be used to understand the biochemical and physiological processes that occur across and within this tissue.
The freshwater fish gill is a multifunctional organ and is the site of gas exchange, osmoregulation, trace metal transport, nitrogenous waste excretion and xenobiotic uptake. Fish are commonly used as indicator species to identify environmental hazards and, as their gills are constantly in contact with water, the gill epithelium is a focal point for studies that seek to understand deleterious effects of environmental toxicants. Whole animal studies can use millions of fish worldwide each year, and efforts are focusing on refining, reducing and replacing (3Rs) these numbers. FiGCS offers an alternative method to study fish toxicology in line with the 3Rs to reduce the numbers of animals in scientific research.
The gills from a freshwater rainbow trout are cleaned and trypsinised resulting in a solution of gill cells in cell culture medium supplemented with FBS and antibiotics. These cells are seeded onto permeable membrane inserts, placed in companion wells and left over night, during which the cells attach. The next day, gill cells are obtained from another rainbow trout in the same way, and these are then seeded on top of the previous days cells. This allows the attachment of mitochondria rich cells between the pavement cells seeded on the first day, and the cells from these two fish can produce between 40 and 72 gill epithelia in culture. Over the next few days the epithelia grow and develop through the assembly of tight junctions between cells. This can be monitored through measurements of transepithelial electrical resistance (TER).