In vitro culture for long-term maintenance of embryonic insect cells from the house cricket Acheta domesticus.

Insect cell culture has become foundational for many areas of research and biotechnological innovation, yet major taxonomic gaps persist, particularly among orthopteran species. Here, we report the isolation and long-term maintenance of primary embryonic cells derived from the house cricket, Acheta domesticus. By isolating cells from late-stage embryos and culturing them at 29°C in Shields and Sang M3 insect cell media supplemented with 1 mg/mL yeast extract, 2.5 mg/mL Bacto Peptone, 100 µg/mL primocin, and 20% heat inactivated FBS, we were able to establish robust primary cultures of small, rounded cells. Passaging via mechanical sloughing preserved cell viability and supported 37.7 cumulative doublings over 287 days, thus, an average doubling time of 182.4 h. The results highlight the importance of conditioned media for sustaining proliferation, while also identifying challenges related to metabolite accumulation and slow doubling times. These preliminary findings provide an avenue for expanding the diversity of insect cell lines, with promising implications for both cellular agriculture and fundamental biotechnology research.