Establishment of a primary culture of haemocytes from the Australian red claw crayfish, Cherax quadricarinatus

In this study, the conditions for establishment of a primary cell culture of haemocytes from the Australian red claw crayfish, Cherax quadricarinatus, were investigated. Five commercial media were assessed for haemocyte viability: Leibovitz L-15 medium (L-15), modified Leibovitz L-15 medium (ML-15, 2 x concentration), Grace's insect medium, DMEM medium, and I-max medium. In addition, L-15 and Grace's insect medium supplemented with 10%, 15%, and 20% charcoal-dextran-treated foetal bovine serum (FBS) were assessed for their effects on haemocyte growth. The optimal culture system was established as follows: L-15 basal medium with the addition of 0.4 g/l glucose, 2 g/l NaCl, 10% FBS, 100 U ml(-1) penicillin, 100 mu g ml(-1) streptomycin, and pH 7.2-7.4. In addition, Giemsa staining of the cultured haemocyte monolayer revealed that two types of cell could be observed: semigranular cells (SGC) and granular cells (GC). Our study could enable further investigation of the immune function in this species.