Over-expression of potato virus X TGBp1 movement protein in transgenic tobacco plants causes developmental and metabolic alterations.

Transgenic Nicotiana tabacum plants expressing the TGBp1 movement protein of potato virus X (PVX) were studied to investigate the effects caused by this protein on plant physiology and development. TGBp1 caused consistent reductions of size and weight in different organs of these plants; however shoot-to-root ratios were similar to those of control plants. Transgenic seedlings showed smaller root meristems and calli derived from TGBp1 leaves grew at a slower rate through successive subcultures. Microscopic observations of TGBp1 plants revealed flattened chloroplasts containing plastoglobuli-like bodies. Further analyses showed a considerable reduction in photosynthetic rate, lower starch levels in leaves and roots, higher nitrate accumulation in leaves and induction of pathogenesis-related (PR) protein genes. Since these changes were not observed when other PVX sequences were expressed in tobacco, we postulate that TGBp1 is an important symptom contributor in PVX infections.

Long-term culture of human embryonic stem cells in feeder-free conditions.

We have demonstrated previously that human embryonic stem (hES) cells possess a characteristic morphologic, antigenic, and molecular profile that can be used to assess the state of ES cells (Carpenter et al., [2004] Dev Dyn 229:243-258). In this manuscript, we have examined the long-term stability of three hES cell lines in feeder-free culture. We demonstrate that the expression of antigens and transcription factors, telomerase activity, telomere length, and karyotype appear stable for all three hES cell lines after continuous culture for over 1 yr. All three lines retained pluripotent differentiation in vitro and in vivo. Although hES cell lines were remarkably stable over the period of analysis, a detailed quantitative analysis of antigen expression by flow cytometry and gene expression by microarray suggested that cell lines show subtle differences in the expression of small subsets of genes upon long-term culture.

Properties of four human embryonic stem cell lines maintained in a feeder-free culture system.

Several laboratories have begun evaluating human ES (hES) cell lines; however, direct comparisons between different hES cell lines have not been performed. We have characterized the properties of four human cell lines maintained in feeder-free culture conditions. Quantitative assessment of surface markers, microarray analysis of gene expression patterns, expression of SOX-2, UTF-1, Rex-1, OCT3/4, CRIPTO, and telomerase activity demonstrated similar patterns in all hES cell lines examined. Undifferentiated hES cells do not respond to neurotransmitters such as acetylcholine, glutamate, and gamma-aminobutyric acid. In addition, the undifferentiated hES cells possess gap junctions. Although similarities in marker expression were observed, allotyping showed that all four lines have a distinct HLA profile, predicting differences in transplantation responses. These data provide the first detailed comparison of different hES cell lines and demonstrate remarkable similarities among lines maintained in identical culture conditions.