Primary cell culture from embryos of the common house spider Parasteatoda tepidariorum

Background Spiders have emerged as valuable models in evolutionary developmental biology, but primary cell cultures from spider embryonic tissues have not been fully explored. In this study, we describe the first successful long-term cultivation of embryonic cells from the common house spider, Parasteatoda tepidariorum . We initiated five independent primary cultures using mechanical and enzymatic dissociation methods, comparing two culture media, Leibovitz’s L-15 and Grace’s Insect Medium, under varying pH conditions. Results Cultures exhibited diverse cell morphologies, including round cells in suspension and elongated, neuron-like cells. The most successful culture, initiated with Grace’s medium at pH 7, was passaged four times and maintained for over six months. We also tested collagen type I-coated wells to improve cell adhesion. Our results indicate that P. tepidariorum embryonic cells proliferate better at pH 7, and Grace’s medium supports long-term growth, while L-15 promotes more cell differentiation. Conclusions This culture system provides a valuable platform for functional genomics studies, with potential applications in evolutionary and developmental biology research.