Ratnayake Dhanushika, Currie Peter D
Australian Regenerative Medicine Institute, Monash University, Clayton, VIC, Australia.
European Molecular Biology Laboratory Australia Melbourne Node, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2019;1889:245-254. doi: 10.1007/978-1-4939-8897-6_14.
This chapter describes a protocol for the isolation of larval zebrafish muscle stem/progenitor cells by fluorescence-activated cell sorting (FACS). This method has been successfully applied to isolate pax3a expressing cells 3 days following needle stab skeletal muscle injury. The cell sorting strategy described here can easily be adapted to any cell type at embryonic or larval stages. RNA extracted from the sorted cells can be used for subsequent downstream applications such as quantitative PCR (qPCR), microarrays, or next generation sequencing.
本章描述了一种通过荧光激活细胞分选（FACS）分离斑马鱼幼体肌肉干细胞/祖细胞的方法。该方法已成功应用于针刺骨骼肌损伤3天后分离表达pax3a的细胞。此处描述的细胞分选策略可轻松适用于胚胎或幼体阶段的任何细胞类型。从分选细胞中提取的RNA可用于后续的下游应用，如定量PCR（qPCR）、微阵列或下一代测序。
