Primary cell culture of XO-SG from red claw crayfish (Cherax quadricarinatus)

In this study, eyestalks of red claw crayfish were dissociated with trypsin to gain single neurons and classified neurons by microscopic inspection. And then, dissociated neurons were cultured into sterile Leibovitz's L-15 medium to screen out proper conditions, etc. The goal of the study is to establish a standard for neurons classification and carry out further research about the endocrine system regulation for crustacean in vitro and in vivo. The results showed that eyestalk XO-SG neurons of red claw crayfish, Cherax quadricarinatus were classified into six different types based on morphological characters with differences on the cell size, and some other differences at microscopic levels. A cell culture approach has been established for XO-SG neurons in vitro, and neurons showed an immediate outgrowth and could regenerate in modified L-15 medium for axon or dendrite. Primary neurons could survive more than 14 d with different growth rate. Some neurons exhibit regeneration of axon or dendrite two days later. Regenerative process can last for 7 days, from then on, the neurons start to shrink and apoptosis. This research provided basis for further study in classifying XO-SG neurons and understanding the endocrine metabolism and regulation at the cell level.