Duressa Tewodros Firdissa, Huybrechts Roger
Insect Physiology and Molecular Ethology Research Group, Division of Animal Physiology and Neurobiology, Biology Department, KU Leuven, Naamsestraat 59, 3000, Leuven, Belgium.
In Vitro Cell Dev Biol Anim. 2016 Jan;52(1):100-6. doi: 10.1007/s11626-015-9952-5. Epub 2015 Oct 1.
Insect cell cultures played central roles in unraveling many insect physiological and immunological processes. Regardless, despite imminent needs, insect cell lines were developed primarily from Dipteran and Lepidopteran orders, leaving many important insects such as Orthopteran locusts under-represented. Besides the lack of cell lines, the slow progress in development of in vitro techniques is attributed to poor communications between different laboratories regarding optimized primary cell cultures. Therefore, we report here about methods developed for primary cell culture of Locusta migratoria hemocyte and phagocytic tissue cells by which we could maintain viable hemocytes in vitro for over 5 d and phagocytic tissue cells for over 12 d. 2-Mercaptoethanol and phenyl-thiourea supplements in Grace's medium together with addition of fetal bovine serum 30 min after cell seeding resulted in a successful setup of the primary cell cultures and a week-long survival of the hemocytes and phagocytic tissue cells in vitro.
昆虫细胞培养在揭示许多昆虫生理和免疫过程中发挥了核心作用。尽管如此，尽管有迫切需求，但昆虫细胞系主要是从双翅目和鳞翅目昆虫中建立起来的，许多重要昆虫，如直翅目蝗虫，在细胞系方面的代表性不足。除了缺乏细胞系外，体外技术发展缓慢还归因于不同实验室之间在优化原代细胞培养方面缺乏沟通。因此，我们在此报告关于开发的飞蝗血细胞和吞噬组织细胞原代细胞培养方法，通过这些方法我们可以在体外使活血细胞维持超过5天，吞噬组织细胞维持超过12天。在格雷斯培养基中添加2-巯基乙醇和苯硫脲，并在细胞接种后30分钟添加胎牛血清，成功建立了原代细胞培养体系，并使血细胞和吞噬组织细胞在体外存活了一周。
