Production of functional proteins: balance of shear stress...

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 21/02 (2006.01) C07K 14/505 (2006.01) C07K 14/705 (2006.01) C12N 5/00 (2006.01) C12N 5/06 (2006.01) C12N 5/08 (2006.01) C12N 9/02 (2006.01) C12N 15/11 (2006.01) C12N 15/67 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2286349

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.

L'invention concerne un procédé relatif à l'élaboration de protéines fonctionnelles, y compris les hormones issues des cellules rénales, qui consiste à établir un processus de co-culture à trois dimensions réagissant à la contrainte tangentielle dans un récipient à parois tournantes. Le mélange naturel des cellules rénales exprime l'enzyme 1-a-hydroxylase, susceptible d'être utilisée pour engendrer la forme active de la vitamine D: 1,25-diOH vitamine D3. Les cultures de fibroblastes et la co-culture de cellules rénales corticales expriment le gène correspondant à l'érythropoïétine et secrètent l'érythropoïétine dans le surnageant de culture. D'autres gènes réagissant à la contrainte tangentielle sont également modulés sous l'effet de ladite contrainte : par exemple, récepteurs de toxines comme la mégaline et la cubuline (gp280). L'invention concerne aussi un procédé conçu pour traiter les individus nécessitant un traitement à base de protéines fonctionnelles, élaborées en processus de co-culture à trois dimensions réagissant à la contrainte tangentielle et établi dans un récipient à parois tournantes.

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