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.