Project description:To investigate transcriptional changes on three different cell lines cultured in 8 different maturation media formulations compared to maintanence media
Project description:We have replaced the right arm of chromosome IX in Saccharomyces cerevisiae with a synthetic version to generate synIXR haploids. The synthetic chromosome features multiple sequeunce modifications. We analyzed gene expression by microarray analysis in three synIXR haploids (1D, 6B, and 22D) to detect any changes in synIXR transcripts or global compensatory changes. Biological triplicates of three synIXR haploids (1D, 6B, and 22D) and corresponding controls (either BY4741 or BY4742) were grown to mid-log phase and collected for mRNA isolation.
Project description:Analysis of 3T3-L1 adipocytes treeated with dexamethasone (Dex) for 6 hours. Dex is a synthetic glucorticoid (GC) receptor agonist. Results provide insight into the effect of glucocorticoids on adipocytes. Differentiated 3T3-L1 adipocytes were cultured in DMEM supplemented with 5% stripped FBS for 24 hours and then treated with 500 nΜ Dex or an equal volume (0.05% v/v of media) of ethanol as a vehicle control for 6 hours at 37°C in DMEM with 190 5% stripped FBS. Total cellular RNA was isolated utilizing the NucleoSpin RNA II kit (Macherey-Nagel).RNA isolates were first quantified by standard spectrophotometry, and then qualitatively evaluated by capillary electrophoresis employing the Bio-Rad Experion system per the manufacturer’s instruction. The final labeled cRNA samples were hybridized overnight to Illumina mouseWG-6 BeadChip arrays, which was performed at UCSF Genomic Core. All treatments were done in triplicates and the same batch of microarrays were used for all treatments. The Illumina expression arrays were pre-processed using lumi package. The differential expression analysis was performed using the Limma package.
Project description:Blood transfusion plays a vital role in modern medicine. However, frequent shortages pose a significant healthcare challenge. Ex vivo manufacturing of red blood cells (RBCs) derived from universal donor pluripotent stem cells is limited by the high cost of recombinant cytokines required for ex vivo erythroid differentiation. Erythropoietin (EPO) signaling through the EPO receptor is indispensable to RBC development, and EPO is one of the most expensive components in erythroid-promoting media. Here, we use design-build-test cycles to develop highly optimized small molecule-inducible synthetic EPO receptors (synEPORs) which are integrated at a variety of genomic loci using homology-directed repair genome editing. We find that integration of synEPOR at the endogenous EPOR locus in an induced pluripotent stem cell producer line cultured with small molecule yields equivalent erythroid differentiation, transcriptomic changes, and hemoglobin production compared to cells cultured with EPO. Due to the dramatically lower cost of small molecules vs. recombinant cytokines, these efforts eliminate one of the most expensive elements of ex vivo culture media—EPO cytokine. As cytokines dependence is a common barrier to ex vivo cell production, these strategies could improve scalable manufacturing of multiple clinically relevant cell types.
Project description:This dataset compares the transcriptome of five laboratory yeast strains (S. cerevisiae) cultured under standardized laboratory consitions (early exponential growth, cultured in synthetic complete glucose media at 30ºC).
Project description:Three human gut microbiome samples from different individuals were cultured in an optimized culture medium with or without the presence of different sugars (10 mM glucose, 20 mM fructose, 10 mM glucose + 20 mM fructose, or 10 mM kestose). Samples were cultured in technical triplicates, and were taken at 0 hr, 1hr, 5 hr, 12 hr and 24 hr of culturing for optical density and metaproteomic analyses. Cultured microbiota cells were subjected to metaproteomics analysis using LC-MS/MS and a TMT approach.
Project description:Analysis of LNCaP cell molecular differences and their response to R1881 in 2D and 3D cultures. Androgen regulated genes were differentially expressed between 2D and 3D cultures. These results provide insights into factors that influence the expression of androgen regulated genes In this study, LNCaP cells cultured in tissue culture plastic (2D) and in the hydrogel (3D) were maintained up to 3 days and 24 days respectively in serum containing media before they were androgen-starved for 48 hours. Cells were either treated with 1nM R1881 or continued to be androgen-deprived (without R1881 with 0.008% ethanol) for another 48 hours prior to cell harvest for gene expression analysis. Biological triplicates were prepared for each condition.
Project description:Transcriptional profiling of U87WT 5 in comparison to U87Mut 5 cells. Three different cultured conditions were tested for 24 h: Serum free media, 50nM IGF1 and 50nM IGF1+ 0.5uM 0SI906
Project description:Yeast lacking the H3 or H4 amino termini, and corresponding wild type strains, were grown in synthetic media. These conditions induce Gcn4-activated transcription. Keywords: Genetic modification