Filice Melissa, Lee C, Mastromonaco Gabriela F
Integrative Biology, University of Guelph, Guelph, Ontario, Canada,
In Vitro Cell Dev Biol Anim. 2014 Oct;50(9):807-10. doi: 10.1007/s11626-014-9790-x. Epub 2014 Jul 2.
The global decline of freshwater fishes has created a need to cryopreserve biological materials from endangered species in an effort to conserve the biodiversity within this taxon. Since maternal gametes and embryos from fish are difficult to cryopreserve, somatic cells obtained from caudal fins have become an increasingly popular resource as they contain both maternal and paternal DNA ensuring valuable traits are not lost from the population. Somatic cells stored in cryobanks can be used to supplement endangered populations with genetically valuable offspring with the use of assisted reproductive technologies. However, initiating primary cell cultures from caudal fin biopsies of endangered species can be challenging as standardized protocols have not yet been developed. The objective of this study was to identify culture conditions, including antibiotic supplementation, biopsy size, and culture temperature, suitable for establishing primary cell cultures of ngege (Oreochromis esculentus), a critically endangered African cichlid. Six-millimeter caudal fin biopsies provided sufficient material to develop a primary cell culture when incubated at 25°C using standard fish cell culture medium containing 1× Primocin. Further investigation and application of these culture conditions for other endangered freshwater fishes is necessary.
全球淡水鱼数量的减少使得有必要对濒危物种的生物材料进行冷冻保存，以努力保护该分类群内的生物多样性。由于鱼类的母本配子和胚胎难以冷冻保存，从尾鳍获取的体细胞已成为越来越受欢迎的资源，因为它们包含母本和父本的DNA，可确保种群中宝贵的性状不会丢失。储存在细胞库中的体细胞可通过辅助生殖技术用于培育具有遗传价值的后代，以补充濒危种群。然而，由于尚未制定标准化方案，从濒危物种的尾鳍活检组织中启动原代细胞培养可能具有挑战性。本研究的目的是确定适合建立极度濒危的非洲丽鱼科鱼类恩格格（Oreochromis esculentus）原代细胞培养的培养条件，包括抗生素添加、活检大小和培养温度。当在25°C下使用含有1× Primocin的标准鱼类细胞培养基孵育时，6毫米的尾鳍活检组织提供了足够的材料来建立原代细胞培养。有必要对这些培养条件进行进一步研究并应用于其他濒危淡水鱼类。
