Ported parallel plate flow chamber and methods for use thereof
A flow chamber and port technology, applied in stress-stimulated microbial growth methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problem of large-scale cultivation without providing multiple flow channel conditions or readout methods Activities and other issues
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[0086] The following examples provide illustrative embodiments. Those skilled in the art, in light of this disclosure and one's ordinary level of skill in the art, will appreciate that the following examples are intended to be illustrative only and that many variations, modifications and changes may be employed without departing from the scope of the disclosed subject matter.
example 1
[0088] Biomarker Analysis of Fluid Flow Regulated Cells
[0089] Differences in gene expression under two different shear stress conditions were tested in human aortic endothelial cells using a flow cell setup.
[0090] Table 1 shows the number of genes changed between human aortic endothelial cells exposed to a wall shear stress of 1.0 Pa for 20 hours compared to cells not exposed to flow. Cells were grown on collagen I-coated growth surfaces under static conditions (no flow) until confluence was reached. Cells were then exposed to fluid flow in the flow chamber at 0.2 or 1.0 Pa for 20 hours, or maintained in static culture for the same amount of time. After 20 hours, use AmbionMIRVANA TM The RNA Isolation Kit (Life Technologies, Foster City, CA, USA) isolated RNA from the cells and performed the analysis on Affymetrix PRIVIEW TM Array (Affymetrix Corporation, Santa Clara, California, USA) for analysis. Three experiments were performed for each condition, repeating the ...
example 2
[0096] Drug treatment of fluid flow preconditioned and statically cultured cells
[0097] PI103(3-[4-(4-morpholino)pyrido[3',2',4,5]furo[3,2-d] Pyrimidin-2-yl]phenol; CAS No. 371935-74-9), a flow cell setup was also used to test for gene expression differences in human aortic endothelial cells under different shear stress conditions.
[0098] Cells were grown on collagen I-coated growth surfaces and treated with 100 nM PI-103 under flow. For the 4 hour time point, cells were exposed to fluid flow for 16 hours and treated with PI-103 for the final 4 hours with flow. Cells were also treated with PI-103 throughout the duration of flow (20 hours). The data were generated using the extracted RNA as described in Example 1 and presented in Affymetrix PRIVIEW TM Array (Affymetrix) was used for analysis. Three experiments were performed for each condition, repeating the microarray analysis.
[0099] Table 2 shows the number of genes that were significantly different between pair...
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