Project description:Examination of the difference in mRNA expression profile between mid-old fibroblasts co-cultured with mid-old fibroblasts and mid-old fibroblasts co-cultured with young fibroblasts
Project description:Investigation of the rejuvenation effect to mid old cell, we co-cultured mid old cell with young cell or mid old cells. After 30days we harvest cells and extrect RNA then anlaysis the gene expression.
Project description:Our research and previous reports showed that fibroblasts could inhibit osteoblast differentiation of Mesenchymal stem cells (MSCs). Thus, RNA-seq was carried to analyze the transcriptome differences among fibroblasts cultured alone and fibroblasts co-cultured with MSCs to find detailed mechanisms about how fibroblasts had functions on MSCs. We then performed gene expression profiling analysis using data obtained from RNA-seq of fibroblasts cultured alone or co-cultured with MSCs.
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:Endothelial cells from 3 WT and 3 Gas6-/- mice were harvested, and were co-cultured with M27 murine lung cancer cells for 48 hours. Control experiments included untreated endothelial cells, and endothelial cells co-cultured with cancer cell media.
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.