Coelomocytes are macrophage-like cells in the body cavity or the coelomic spaces of many invertebrates and play major roles in their physiology and immunology. Their structure, function and diversity, however, is still poorly understood.

Tardigrades are micrometazoans inhabiting a wide variety of environments and with an ability to survive extreme conditions. Coelomocytes (“storage cells”) represent an important part of tardigrade physiology, storing and distributing energy and possibly also having immunological functions. Few studies of tardigrade cell biology have been reported and neither primary nor continuous cell cultures have been established. Tardigrades are normally found and also cultured in an environment rich in microorganisms, some of which may even be of symbiotic value.

In this study we have tried to establish a primary culture of storage cells in the eutardigrade Richtersius coronifer. Different cell media and concentrations of fetal bovine serum (FBS) were tested. Extracting cells from the tardigrades in an antiseptical environment is challenging since it has to be done under a microscope and contamination from the tardigrades surface is also a problem. To avoid this we tried culturing with high concentrations of antibiotics and antimycotics. We managed to keep the cells viable for up to 18 days in Grace insect medium with 10 % FBS at 20-22°C. The medium was changed every third day. 10x Antibiotic-Antimycotic and 5x of Penicillin-Streptomycin were used to minimize contamination. These concentrations reduce the bacterial abundance, but contamination with fungi was still an issue. Cell morphology evaluation was performed daily and no obvious toxic effects on the cells was observed. Cell viability and cell division were evaluated with Trypan blue staining and cell counting in a haemocytometer. The results indicate that the cells are viable and that some cell division occurs, however more studies need to be performed to confirm this. Still, this study provides the first evidence that primary cultures of storage cells from tardigrades are possible to establish, but the culturing method has to be refined to avoid contamination.