Establishment of long-term ostracod epidermal culture
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Establishment of long-term ostracod epidermal culture Siân R. Morgan 1,2 & Laura Paletto 1,2,3 & Benjamin Rumney 1,2,3 & Farhana T. Malik 4,5 & Nick White 1,2,3 & Philip N. Lewis 1,2 & Andrew R. Parker 4,5 & Simon Holden 6 & Keith M. Meek 1,2 & Julie Albon 1,2,3 Received: 5 June 2020 / Accepted: 9 September 2020 / Editor: Tetsuji Okamoto # The Author(s) 2020
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. Keywords Crustacean . Ostracod . Carapace . Epidermal . Culture
Introduction Myodocopid ostracods (or ‘seed shrimps’) are a suborder of small marine crustaceans of length between 1 and 32 mm, which pass through a series of six, juvenile growth stages known as instars before reaching the adult stage (McKenzie et al. 1999). Their shrimp-like bodies are suspended within a relatively thin (20–60 μm), bivalved calcified carapace by ‘central adductor muscles’, which run through the body * Julie Albon [email protected] 1
School of Optometry and Vision Sciences, Cardiff University, Cardiff CF24 4HQ, UK
2
Cardiff Institute for Tissue Engineering and Repair, School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff CF10 3NB, UK
3
Vivat Scientia Bioimaging Laboratories, School of Optometry and Vision Sciences, Cardiff University, Cardiff CF24 4HQ, UK
4
Lifescaped, Somerset House, London WC2R 1LA, UK
5
Green Templeton College, University of Oxford, Woodstock Road, Oxford OX2 6HD, UK
6
Defence Science and Technology Laboratory (DSTL), Porton Down, Salisbury SP4 0JQ, UK
attached to each valve; a hinge joins the two valves enabling them to open and close their carapace while preserving its rigidity (McKenzie et al. 1999). The majority of myodocopid ostracods are benthic; they bury up to 1 cm depth in the sediments of the ocean floor (McKenzie et al. 1999).
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