The techniques for establishing and maintaining invertebrate tissues and cells in culture remain difficult due to the diversity of invertebrates and their structural and physiological characteristics. Research involving invertebrate cell cultures continues to increase, although the number of cell lines used is still limited. This manual gives detailed descriptions of the technical procedures for the establishment of primary invertebrate cell cultures in vitro. Nutritional requirements, culture media, and species-specific methods for both cell and organ cultures as well as useful techniques for studies on cultured cells are described. The Appendix lists established cell lines available for research with information on the composition of their physiological and nutrient solutions. This comprehensive manual, the first of its kind, is a valuable reference for investigators working with invertebrate cell cultures in academia and industry.