Development and utilization of crustacean long-term primary cell cultures: Ecdysteroid effectsin vitro

Abstract

Summary Long-term maintenance of lobster, Homarus americanus and crayfish, Pacifasticus leniusculus primary cell cultures of testicular and hematopoietic tissues, for 11 and 3 months, respectively, succeeded in a modified Medium 199 supplemented with 10% fetal bovine serum (pH 7.5, 200°C). In addition, NaC1 was used to adjust the lobster culture medium to 1000 mOsm and the crayfish medium to 400 mOsm. Proline concentration was also elevated. Testes were dissociated with 200 U/ml type II collagenase 2–3 days prior to culture. Lobster hemocytes reacted to 10−7 M 20-hydroxyecdysone (20-HE) by reducing contact inhibition and increasing invasive behavior one week after hormonal exposure. The presence of 10−7 M 20-HE caused mesodermal cell death and spermatogonial proliferation in lobster testicular cell cultures within one week. Crayfish testicular mesodermal cells formed vacuoles 5 days after exposure to 10−8 M 20-HE. These results are discussed in relation to the cellular events that occur in vivo during pre...