Project description:Brain metastases are highly resistant to chemotherapy. Brain metastases are surrounded and infiltrated by activated astrocytes. To examine the genes whose expression is associated with chemo-resistance of brain-metastasized cancer cells, gene expression data were collected and analyzed from breast cancer cells and lung cancer cells co-cultured with astrocytes. Fibroblast cells were used as control. Human lung cancer cell PC14 was co-cultured with mouse astrocytes or fibroblasts for two rounds. Total RNAs were extracted from co-cultured cells and hybridized to human microarray.
Project description:Brain metastases are highly resistant to chemotherapy. Brain metastases are surrounded and infiltrated by activated astrocytes. To examine the genes whose expression is associated with chemo-resistance of brain-metastasized cancer cells, gene expression data were collected and analyzed from breast cancer cells and lung cancer cells co-cultured with astrocytes. Fibroblast cells were used as control. Total RNAs were extracted from co-cultured cells and hybridized to human microarray and mouse microarrays to detect human cell and mouse cell specific gene expression data.
Project description:Brain metastases are highly resistant to chemotherapy. Brain metastases are surrounded and infiltrated by activated astrocytes. To examine the genes whose expression is associated with chemo-resistance of brain-metastasized cancer cells, gene expression data were collected and analyzed from breast cancer cells and lung cancer cells co-cultured with astrocytes. Fibroblast cells were used as control. Total RNAs were extracted from co-cultured cells and hybridized to human microarray and mouse microarrays to detect human cell and mouse cell specific gene expression data.
Project description:We report the first RNA profiing of mammalian neural cells grown in vitro. About 50 million reads are generated by RNA-seq from rat astrocytes, neurons and oligodendrocyte precursor cells in primary culture. These data are compared with theose generated from the correponding mouse neural cells that are acutely purified from brains. Cross-species RNA-seq data analysis revealed hundreds of genes that are differentially expressed between cultured and acutely purified cells. Astrocytes have more such genes compared to neurons and oligodendrocyte precursor cells, indicating that signalling pathways are greatly perturbed in cultured astrocytes. mRNA profiles of rat astrocytes, neurons and oligodendrocyte precursor cells cultured in vitro
Project description:We report the first RNA profiing of mammalian neural cells grown in vitro. About 50 million reads are generated by RNA-seq from rat astrocytes, neurons and oligodendrocyte precursor cells in primary culture. These data are compared with theose generated from the correponding mouse neural cells that are acutely purified from brains. Cross-species RNA-seq data analysis revealed hundreds of genes that are differentially expressed between cultured and acutely purified cells. Astrocytes have more such genes compared to neurons and oligodendrocyte precursor cells, indicating that signalling pathways are greatly perturbed in cultured astrocytes.
Project description:Molecular mechanisms of the cancer cells-carcinoma associated fibroblasts (CAF) interactions growing in vitro conditions as a co-culture. The five canine mammary cancer cell lines were cultured with CAF (isolated from canine mammary cancer) for 72hrs. Then, the cancer cells and CAFs were sorted and the gene expression analysis was conducted. The control for each co-cultured cell line was the same cell line growing as a single culture.
Project description:Profiling of transcriptional changes in rat astrocytes when co-cultured with neurons: comparison of astrocytes cultured alone with astrocytes co-cultured with mouse hippocampal neurons. Co-cultured astrocytes are isolated using cold jet, a novel tool for these neuron-glia cultures. Over the last decade, the importance of astrocyte-neuron communication in neuronal development and synaptic plasticity has become increasingly clear. Since neuron-astrocyte interactions represent highly dynamic and reciprocal processes, we hypothesized that at least part of the involved astrocyte genes may be regulated as a consequence of their interactions with maturing neurons. In order to identify such neuron-induced astrocyte genes in vitro, we tested the effectiveness of the ‘cold jet’, a new method for separation of neurons from co-cultured astrocytes. The cold jet method is performed under ice-cold conditions and avoids protease-mediated isolation of astrocytes or time-consuming centrifugation, yielding intact astrocyte mRNA with approximately 90% of neuronal RNA removed. Using this method, we executed genome-wide profiling in which RNA derived from astrocyte-only cultures was compared with astrocyte RNA derived from differentiating neuron-astrocyte co-cultures. Data analysis revealed changes in expression of a large number of mRNAs and biological processes, including novel findings. Thus, cold jet is an efficient method to separate astrocytes from neurons in co-culture, and in this study reveals that neurons induce robust gene-expression changes in co-cultured astrocytes.