High-throughput transformation and cryopreservation of Arabidopsis suspension-cultured T87 cells for functional genomics

Abstract

We are currently analyzing Arabidopsis metabolism-related genes with a combination of transgenic suspension-cultured T87 cells and transcriptome/metabolome analyses. Here we established a high-troughput protocol of Agrobacterium-mediated transformation of the T87 cells. In our routine procedure, a person works five days for 24 Agrobacterium strains carrying individual manipulated genes for transformation of plant cells and picks more than ten transgenic calli two or three weeks later. Moreover, we developed a new protocol for cryopreservation of transgenic T87 cell lines, by which a person can handle 200 vials with transgenic suspension-cultured cells and stock them in liquid nitrogen within a single day. The protocols we developed would be crucial to high-throughput analyses, especially of the metabolisms observed in the T87 cells, and could be applicable for other plant suspension-cultured cell lines.