As insect cell lines can be used in baculovirus expression systems for producing recombinant proteins, there is a need to establish new cell lines. Insect cell lines are thought to be useful in production of humanized recombinant proteins. Hence, we aimed to establish a culture from larval ovaries of Bombyx mori to investigate the cell characteristics and growth properties. Two sets of primary cell cultures were established from ovaries of fifth instar larvae. Characterization of cell line was carried out by comparing the PCR products of cell line and silkworms from COI (cytochrome c oxidase subunit I) specific primers. In the first set of cultures, the culture population was found to be heterogeneous and cell sizes were variable. Fibroblastic, epithelial-like, spindle-like, and round cells were observed after 7th day. Interestingly, contractions were observed in muscle-like cells at the end of first month, and stayed stable until the fourth month. At the beginning of the fourth month, because cell death increased significantly, cell concentrations decreased. In the second set of cultures, it was determined that doubling time was about 72 h with high viability and confluency of cells were higher in early passages. PCR which was conducted for characterization of cells resulted in a 710 bp product in each cell line and silkworms. In conclusion, growth rate detected in this study was observed to be compatible with literature. Also, it was thought that if primary culture is supported with growth stimulators, growth rate can be more accelerated and this primary culture can be used more efficiently in recombinant protein production studies.