United States of America As Represented by the Administrator of Nasa

The United States Government As Represented by the Department of Veterans Affairs

The Administrators of the Tulane Educational Fund

Tulane University

Timothy Grant Hammond

Thomas John Goodwin

James Howard Kaysen

Cheryl Anne Nickerson

Pierre J Verroust

Patricia L Allen

A method for the production of functional proteins including hormones by renal cells in a three dimensional culturing process responsive to shear stress uses a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-&#x3b1;-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating an in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel.

Production of functional proteins: balance of shear stress and gravity

Embodiments of a method for the production of human urokinase are disclosed. Also disclosed are embodiments of a cell culture well-suited for use with the disclosed method. The method involves culturing urokinase-producing cells, such as immortalized human renal cells, in a cell culture. The cell culture comprises microcarrier structures and a tissue culture medium. The urokinase production is allowed to occur while the cell culture remains relatively static, i.e., the cell culture is not substantially mixed.

Production of urokinase in a three-dimensional cell culture

The present invention provides methods for utilizing a form of optimized suspension culture to examine the infectivity of pathogenic organisms and agents in human cells and tissues. Also provided are methods using a rotating wall vessel to predict chemosensitivity of cells and tissues to toxins and chemotherapeutic agents. These culture conditions potentiate spatial colocalization and three-dimensional assembly of individual cells into large aggregates which more closely resemble the in vivo tissue equivalent. In this environment, dissociated cells can assemble and differentiate into macroscopic tissue aggregates several millimeters in size. These culture conditions allow for better cellular differentiation and formation of three-dimensional cellular aggregates, more efficient cell-to-cell interactions, the in in vivo-like exchange of growth factors and greater molecular scaffolding facilitating mechanical stability for cells. The suspension culture system offers a new approach for studying microbial infectivity from the perspective of the host-pathogen interaction and also for analyzing chemosensitivity to toxins and chemotherapeutic agents.

Methods for modeling infectious disease and chemosensitivity in cultured cells and tissues

The present invention provides a method for production of functional proteins including hormones by renal cells in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-a-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel.

The present invention provides for a method of culturing cells and inducing the expression of at least one gene in the cell culture. The method provides for contacting the cell with a transcription factor decoy oligonucleotide sequence directed against a nucleotide sequence encoding a shear stress response element.

The present invention provides novel renal receptors for ligands. Cubilin and megalin are representative examples of such renal receptors. Also provided are potential uses of these renal receptors for treating toxicity in various tissues and for detecting renal damage.

Growing community of inventors

New Orleans, LA, United States of America

Timothy Grant Hammond