Project description:Rotating wall vessel (RWV) grown A549s compared to monolayer grown A549s infected with F. tularensis SchuS4 over a 22 hour time-course (2, 6 and 22 h post infection)
Project description:A549 epithelial cells grown under two different conditions were compared. A549s grown in rotating wall vessel cultures disp[lay a more in-vivo like phenotype. The data here describe the transcriptional differences between the two culture methods
Project description:A549 epithelial cells grown under two different conditions were compared. A549s grown in rotating wall vessel cultures display a more in-vivo like phenotype. The data here describe the transcriptional differences between the two culture methods when infected with Francisella tularensis SchuS4 over a 22 hour time-course
Project description:A549 epithelial cells grown under two different conditions were compared. A549s grown in rotating wall vessel cultures disp[lay a more in-vivo like phenotype. The data here describe the transcriptional differences between the two culture methods 1 colour; Cy3 labelled, 5 biological replicates for each culture condition, A549s grown in RWVs (RWV) compared to A549 grown in monolayers (mono)
Project description:This study explores the connection between changes in gene expression and the genes that determine strain survival during suspension culture, using the model eukaryotic organism, Saccharomyces cerevisiae. The Saccharomyces cerevisiae homozygous diploid deletion pool, and the BY4743 parental strain were grown for 18 hours in a rotating wall vessel, a suspension culture device optimized to minimize the delivered shear. In addition to the reduced shear conditions, the rotating wall vessels were also placed in a static position or in a shaker in order to change the amount of shear stress on the cells. Keywords: shear stress, time course
Project description:A549 epithelial cells grown under two different conditions were compared. A549s grown in rotating wall vessel cultures display a more in-vivo like phenotype. The data here describe the transcriptional differences between the two culture methods when infected with Francisella tularensis SchuS4 over a 22 hour time-course 1 colour; Cy3 labelled, 6 biological replicates for each culture condition and time-point, A549s grown in RWVs (RWV) compared to A549 grown in monolayers (mono), Infected vs. NaM-CM-/ve
Project description:Full title: Three-dimensional culture of AIDS-NHL cells influences gene expression related to B-cell development, proliferation and survival The AIDS-NHL-derived cell line, UMCL01-101, representing diffuse large B-cell lymphoma of immunoblastic morphology (AIDS-IBL), was grown in conventional, static suspension culture or three-dimensionally (3D) in the Rotating Wall Vessel (RWV) bioreactor. The objective was to assess the impact on gene expression of growth as a three-dimensional tissue assembly. Global gene expression analysis was performed on UMCL01-101 cells grown under either condition using Affymetrix microarray. UMCL01-101 cells were cultured in the Rotating Wall Vessel bioreactor to form 3D assemblies, or in conventional suspension culture, for 15 days. RNA was prepared from triplicate samples under each growth condition and submitted for microarray analysis.
Project description:Full title: Three-dimensional culture of AIDS-NHL cells influences gene expression related to B-cell development, proliferation and survival The AIDS-NHL-derived cell line, UMCL01-101, representing diffuse large B-cell lymphoma of immunoblastic morphology (AIDS-IBL), was grown in conventional, static suspension culture or three-dimensionally (3D) in the Rotating Wall Vessel (RWV) bioreactor. The objective was to assess the impact on gene expression of growth as a three-dimensional tissue assembly. Global gene expression analysis was performed on UMCL01-101 cells grown under either condition using Affymetrix microarray.
Project description:Investigate the functional capabilities of human iPSC-derived liver organoids generated on Matrigel or self-assembed in rotating wall vessel (RWV) via bulk RNA-seq, RT-qPCR and immunostaining, to provide a simple and high-throughput way to generate Matrigel-free liver organoids for research and clinical applications
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.
