Department of Pediatrics (Pediatric Hematology/Oncology and Cellular Therapy) and Department of Developmental and Molecular Biology, Albert Einstein College of Medicine and Montefiore Medical Center, Bronx, NY, USA.
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2023;2567:233-249. doi: 10.1007/978-1-0716-2679-5_15.
The zebrafish as a model organism is well known for its versatile genetics, rapid development, and straightforward live imaging. It is an excellent model to study hematopoiesis because of its highly conserved ontogeny and gene regulatory networks. Recently developed highly specific transgenic reporter lines have allowed direct imaging and tracking of hematopoietic stem and progenitor cells (HSPCs) in live zebrafish. These reporter lines can also be used for fluorescence-activated cell sorting (FACS) of HSPCs. Similar to mammalian models, HSPCs can be transplanted to reconstitute the entire hematopoietic system of zebrafish recipients. However, the zebrafish provides unique advantages to study HSPC biology, such as transplants into embryos and high-throughput chemical screening. This chapter will outline the methods needed to identify, isolate, and transplant HSPCs in zebrafish.
斑马鱼作为一种模式生物,以其多样的遗传学、快速的发育和简单的活体成像而闻名。由于其高度保守的个体发生和基因调控网络,它是研究造血的优秀模型。最近开发的高度特异性转基因报告线允许对活体斑马鱼中的造血干细胞和祖细胞 (HSPC) 进行直接成像和跟踪。这些报告线也可用于 HSPC 的荧光激活细胞分选 (FACS)。与哺乳动物模型类似,HSPC 可以被移植以重建斑马鱼受者的整个造血系统。然而,斑马鱼为研究 HSPC 生物学提供了独特的优势,例如移植到胚胎中和高通量化学筛选。本章将概述在斑马鱼中鉴定、分离和移植 HSPC 所需的方法。