He Fangfang, Tu Lingge, Chan Leslie, Leung Anskar, Sun Xuan
Department of Medicine, School of Clinical Medicine, LKS Faculty of Medicine, The University of Hong Kong.
Department of Medicine, School of Clinical Medicine, LKS Faculty of Medicine, The University of Hong Kong; ZeBlast Technology Limited, Hong Kong Science Park.
J Vis Exp. 2024 Dec 20(214). doi: 10.3791/67463.
Intravenous (IV) injection is widely recognized as the most effective and commonly utilized method for achieving systemic delivery of substances in mammalian research models. However, its application in adult zebrafish for drug delivery, stem cell transplantation, and regenerative and cancer studies has been limited due to the challenges posed by their small body size and intricate blood vessels. To overcome these limitations, alternative injection techniques such as intracardiac and retro-orbital (RO) injection have been explored in the past for stem cell transplantation in adult zebrafish. However, these techniques have their drawbacks, including the need for meticulous injection techniques or increased risk of mortality. In this study, we have developed a refined and optimized IV injection procedure specifically tailored to adult zebrafish, addressing the challenges associated with their unique anatomy. To demonstrate the effectiveness of this technique, we performed successful IV injections of whole kidney marrow cells from Tg(mpo: EGFP) fish and FITC-dextran dye into adult Casper fish. The subsequent visualization of injected cells and dyes using a fluorescence microscope confirmed their successful delivery and engraftment within the zebrafish. Furthermore, we demonstrated that compared with the intracardiac and RO injections, the IV injection resulted in improved survival rates and engraftment efficiency in treated zebrafish. This approach enables precise delivery and localization of substances and holds great potential for large-scale drug and chemical screening using adult zebrafish. Additionally, the ability to visually track the injected cells and dyes provides invaluable insights into their engraftment, migration, and interactions with host tissues, enabling a more comprehensive evaluation of therapeutic effects and biological processes in zebrafish models.
静脉注射被广泛认为是在哺乳动物研究模型中实现物质全身递送的最有效且最常用的方法。然而，由于成年斑马鱼体型小且血管复杂带来的挑战，其在成年斑马鱼用于药物递送、干细胞移植以及再生和癌症研究中的应用一直受到限制。为克服这些限制，过去曾探索过替代注射技术，如心内注射和眶后（RO）注射，用于成年斑马鱼的干细胞移植。然而，这些技术有其缺点，包括需要精细的注射技术或增加死亡风险。在本研究中，我们开发了一种专门针对成年斑马鱼优化的精细静脉注射程序，解决了与其独特解剖结构相关的挑战。为证明该技术的有效性，我们成功地将来自Tg(mpo:EGFP)鱼的全骨髓细胞和FITC-葡聚糖染料静脉注射到成年Casper鱼体内。随后使用荧光显微镜对注射的细胞和染料进行可视化，证实它们成功递送至斑马鱼体内并实现了植入。此外，我们证明与心内注射和眶后注射相比，静脉注射提高了处理后斑马鱼的存活率和植入效率。这种方法能够精确递送和定位物质，在使用成年斑马鱼进行大规模药物和化学筛选方面具有巨大潜力。此外，可视化追踪注射的细胞和染料的能力为它们的植入、迁移以及与宿主组织的相互作用提供了宝贵的见解，从而能够更全面地评估斑马鱼模型中的治疗效果和生物学过程。
