Metabolic Fingerprinting and Profiling of Arabidopsis thaliana Leaf and its Cultured Cells T87 by GC/MS

Cell suspension cultures are now recognized as important model materials for plant bioscience and biotechnology. Very few studies of metabolic comparisons between cell cultures and original plants have been reported, even though the biological identity of cultured cells with the normally grown plant is of great importance. In this study, a comparison of the metabolome for primary metabolites extracted from the leaves of Arabidopsis thaliana and cultured cells from an Arabidopsis suspension culture (cell line T87) was performed. The results suggest that although cell suspension cultures and Arabidopsis leaves showed similarities in the common primary metabolite profile, nonetheless, moderate differences in quantitative profile were revealed.

Acetonitrile is better than ethanol as a dehydrating agent for cells prepared for SEM

Acetonitrile has been shown to be a better dehydrating agent than ethanol for whole tissue prepared for TEM studies. In this paper we show that it is also a better choice for dehydration of cell cultures prepared for SEM studies. The procedures used for the isolation and culture of cardiac cells in the present study have been outlined in detail by Schroedl and Hartzell (1983) and Freerksen, et al. (1984). Cells were cultured in suspension using HARV-Bioreactors (Synthecon, Inc., Friendswood, TX) for 24 hours to allow attachment and then fed each subsequent 48 hours. After a total of six days in suspension culture, cell/bead clusters were collected and washed with PBS.Primary fixation was done at 4°C, all other steps were performed at room temperature. The buffer used in all cases was 0.1M cacodylate, pH 7.3. The cell/bead cultures were processed according to the following protocol. All of the beads were placed in a 1.5 ml microfuge tube and fixed in 2% buffered glutaraldehyde overnight followed by two 5 min washes in buffer, postfixation in 1% buffered OsO4 for 90 min and three washes in buffer. One half of the beads were then placed in another microfuge tube and dehydrated in acetonitrile, while the remaining beads were dehydrated in ethanol.