Abstract
The rotating wall vessel has gained popularity as a clinical cell culture tool to produce hormonal implants. It is desirable to understand the mechanisms by which the rotating wall vessel induces genetic changes, if we are to prolong the useful life of implants. During rotating wall vessel culture gravity is balanced by equal and opposite hydrodynamic forces including shear stress. The current study provides the first evidence that shear stress response elements, which modulate gene expression in endothelial cells, are also active in epithelial cells. Rotating wall culture of renal cells changes expression of select gene products including the giant glycoprotein scavenger receptors cubulin and megalin, the structural microvillar protein villin, and classic shear stress response genes ICAM, VCAM and MnSOD. Using a putative endothelial cell shear stress response element binding site as a decoy, we demonstrate the role of this sequence in the regulation of selected genes in epithelial cells. However, many of the changes observed in the rotating wall vessel are independent of this response element. It remains to define other genetic response elements modulated during rotating wall vessel culture, including the role of hemodynamics characterized by 3-dimensionality, low shear and turbulence, and cospatial relation of dissimilar cell types.
Publication types
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Comparative Study
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Carrier Proteins / biosynthesis
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Carrier Proteins / genetics
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Cell Adhesion Molecules / biosynthesis
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Cell Adhesion Molecules / genetics
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Cell Count
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Cell Culture Techniques / instrumentation
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Cell Culture Techniques / methods*
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Cell Differentiation
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Endosomes / metabolism
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Epithelial Cells / cytology
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Epithelial Cells / metabolism
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Gene Expression Regulation*
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Gravitation
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Heat-Shock Proteins / biosynthesis
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Heat-Shock Proteins / genetics
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Heymann Nephritis Antigenic Complex
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Humans
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Kidney Cortex / cytology*
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Kidney Cortex / metabolism
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Kidney Tubules, Proximal / cytology
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Kidney Tubules, Proximal / metabolism
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Membrane Glycoproteins / biosynthesis*
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Membrane Glycoproteins / genetics
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Microfilament Proteins / biosynthesis
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Microfilament Proteins / genetics
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Oligonucleotides, Antisense / pharmacology
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Prostheses and Implants
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Rats
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Rats, Sprague-Dawley
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Receptors, Cell Surface / biosynthesis*
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Receptors, Cell Surface / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Rotation
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Stress, Mechanical*
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Superoxide Dismutase / biosynthesis
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Superoxide Dismutase / genetics
Substances
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Carrier Proteins
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Cell Adhesion Molecules
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Heat-Shock Proteins
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Heymann Nephritis Antigenic Complex
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Membrane Glycoproteins
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Microfilament Proteins
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Oligonucleotides, Antisense
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Receptors, Cell Surface
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intrinsic factor-cobalamin receptor
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villin
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Superoxide Dismutase