摘要
Maintaining chromosome euploidy in zebrafish embryonic cells is challenging because of the degradation of genomic integrity during cell passaging.In this study,we report the derivation of zebrafish cell lines from single blastomeres.These cell lines have a stable chromosome status attributed to BMP4 and exhibit continuous proliferation in vitro.Twenty zebrafish cell lines are successfully established from single blastomeres.Single-cell transcriptome sequencing analysis confirms the fidelity of gene expression profiles throughout long-term culturing of at least 45 passages.The long-term cultured cells are specialized into epithelial cells,exhibiting similar expression patterns validated by integrative transcriptomic analysis.Overall,this work provides a protocol for establishing zebrafish cell lines from single blastomeres,which can serve as valuable tools for in vitro investigations of epithelial cell dynamics in terms of lifeedeath balance and cell fate determination during normal homeostasis.
基金
supported by the National Key Research and Development Program of China(2018YFA0801003 and 2019YFA0109900)
the Center for Excellence in Molecular Cell Science(2021DF06)
supported by the Shanghai Municipal Science and Technology Major Project.
