School of Optometry and Vision Sciences, Cardiff University, Cardiff, CF24 4HQ, UK.
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.
In Vitro Cell Dev Biol Anim. 2020 Oct;56(9):760-772. doi: 10.1007/s11626-020-00508-8. Epub 2020 Oct 9.
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.
原生甲壳动物细胞培养始于 20 世纪 60 年代，但迄今为止，仅建立了有限的细胞系。Lerneriopsis skogsbergi 是一种隶属于涟虫目的小型等足目甲壳动物，其身体被一层薄而坚韧的透明双瓣甲壳所包裹，眼睛可以透过甲壳看到内部。表皮层覆盖在甲壳的内表面，负责甲壳的合成。本研究旨在建立 Lerneriopsis skogsbergi 的体外表皮组织和细胞培养方法。首先，确定了维持这种表皮组织活力和细胞增殖能力的最佳环境。接下来，建立了用于组织培养的表皮层显微解剖技术，最后，确定了表皮细胞的分离方法。在整个过程中，保持无菌、细胞活力和增殖是关键。这种新型的 Lerneriopsis skogsbergi 表皮组织和细胞培养方法，增加了我们对甲壳动物细胞生物学和甲壳动物复杂生物合成的理解。此外，这些技术在生物材料制造、军事和渔业等领域具有很大的潜力，例如体外毒性测试。
