Establishment of long-term ostracod epidermal culture

Abstract

Primary crustacean cell culture was introduced in the 1960s, but to date limited cell lines have been established. Skogsbergia lerneri is a myodocopid ostracod, which has a body enclosed within a thin, durable, transparent bivalved carapace, through which the eye can see. The epidermal layer lines the inner surface of the carapace and is responsible for carapace synthesis. The purpose of the present study was to develop an in vitro epidermal tissue and cell culture method for S. lerneri. First, an optimal environment for the viability of this epidermal tissue was ascertained, while maintaining its cell proliferative capacity. Next, a microdissection technique to remove the epidermal layer for explant culture was established and finally, a cell dissociation method for epidermal cell culture was determined. Maintenance of sterility, cell viability and proliferation were key throughout these processes. This novel approach for viable S. lerneri epidermal tissue and cell culture augments our understanding of crustacean cell biology and the complex biosynthesis of the ostracod carapace. In addition, these techniques have great potential in the fields of biomaterial manufacture, the military and fisheries, for example, in vitro toxicity testing.

The ultrastructural development and 3D reconstruction of the transparent carapace of the ostracod Skogsbergia lerneri

“…Skogsbergia lerneri were collected from the east coast of the Florida Keys following the granting of Special Activity licenses (SAL-16-1796-SR and SAL-19-1796-SR) and Florida Keys National Marine Sanctuary permits (FKNMS-2016-116 and FKNMS-2018-116). They were transported to Cardiff University and maintained in purpose-built aquaria under controlled aquaculture conditions as described in detail previously (Morgan et al 2020 ). Each ostracod was identified with respect to its gender and developmental stage via its shape, length, height and number of furcal claws, as categorised previously by Cohen ( 1983 ) (see Table 1 ).…”

Stem Cells and Innate Immunity in Aquatic Invertebrates: Bridging Two Seemingly Disparate Disciplines for New Discoveries in Biology

“…In crustaceans, long-term primary cultures of crayfish HPT cells were possible with the use of astakine, a new invertebrate cytokine directly involved in hematopoiesis, thus allowing the study of the molecular mechanism involved in the production of hemocytes ( 84 , 252 , 274 ). Transfection methods were also studied on the hematopoietic primary culture system from Cherax quadricarinatus ( 275 ) and efforts were invested in long-term ostracod epidermal cultures ( 276 ).…”