Department of Ecology, Environment and Plant Sciences (DEEP), Stockholm University, Stockholm, Sweden.
Umeå Plant Science Centre (UPSC), Department of Plant Physiology, Umeå University, Umeå, Sweden.
Methods Mol Biol. 2024;2722:171-200. doi: 10.1007/978-1-0716-3477-6_13.
Inducing the differentiation of specific cell type(s) synchronously and on-demand is a great experimental system to understand the sequential progression of the cellular processes, their timing and their resulting properties for distinct isolated plant cells independently of their tissue context. The inducible differentiation in cell suspension cultures, moreover, enables to obtain large quantities of distinct cell types at specific development stage, which is not possible when using whole plants. The differentiation of tracheary elements (TEs) - the cell type responsible for the hydro-mineral sap conduction and skeletal support of plants in xylem tissues - has been the most studied using inducible cell suspension cultures. We herein describe how to establish and use inducible pluripotent suspension cell cultures (iPSCs) in Arabidopsis thaliana to trigger on-demand different cell types, such as TEs or mesophyll cells. We, moreover, describe the methods to establish, monitor, and modify the sequence, duration, and properties of differentiated cells using iPSCs.
同步且按需诱导特定细胞类型的分化是一个很好的实验系统,可以帮助我们了解细胞过程的顺序进展、它们的时间以及它们为独立于组织背景的特定分离植物细胞带来的特性。悬浮细胞培养中的诱导分化,此外,还能够在特定的发育阶段获得大量不同的细胞类型,这在使用整个植物时是不可能的。使用诱导悬浮细胞培养,对负责木质部组织中水力矿物汁液传导和骨骼支撑的细胞类型——导管分子 (TEs) 的分化进行了最深入的研究。本文描述了如何在拟南芥中建立和使用可诱导的多能悬浮细胞培养物 (iPSC) 来按需触发 TEs 或叶肉细胞等不同的细胞类型。此外,我们还描述了使用 iPSC 建立、监测和修改分化细胞的序列、持续时间和特性的方法。