Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5800811

Title:Artificial skin prepared from coclagen matrix containing transforming growth factor-.beta. having a collagen binding site
Inventors:Hall, Frederick L.; Glendale, CA, USA
Nimni, Marcel E.; Santa Monica, CA, USA
Tuan, Tai-Lan; Anaheim, CA, USA
Wu, Lingtau; Arcadia, CA, USA
Cheung, David T.; Arcadia, CA, USA
Summary:This invention introduces a method of preparing artificial skin from a collagen matrix containing a transforming growth factor-beta fusion protein. Described herein are methods of impregnating a collagen matrix with a transforming growth factor-? having a collagen-binding site to bind the growth factor to the collagen matrix, which is followed by a process of incubating the impregnated matrix with a source of fibroblasts and mesenchymal stem cells to form a captured population of mesenchymal stem cells within the impregnated matrix. Further disclosed are methods by which incubation of the resultant matrix with a source of keratinocytes results in the epithelialization of the matrix to form an artificial skin. Methods, factors and conditions are elucidated, as are therapeutic applications in the reduction of recovery time after surgery, and in the replacement of skin damaged by disease or injury.
Abstract:An artificial skin is prepared by impregnating a collagen matrix with a transforming growth factor-.beta. having a collagen-binding site to bind the growth factor to the collagen matrix, incubating the impregnated matrix with a source of fibroblasts and mesenchymal stem cells to form a captured population of mesenchymal stem cells within the impregnated matrix and incubating the resultant matrix with a source of keratinocytes which epithelialize the matrix to form an artificial skin. The collagen matrix is preferably in the form of a collagen sheet. The transforming growth factor-.beta. can be transforming growth factor-.beta..sub.1, transforming growth factor-.beta..sub.2 or transforming growth factor-.beta..sub.3. Preferably, the transforming growth factor-.beta. having a collagen binding site is a fusion protein having a purification tag, at least one proteinase site, an extracellular matrix binding site and a transforming growth factor active fragment. The extracellular matrix binding site binds collagen, fibronectin or a cell surface. A method of preparing the fusion protein involves purifying and renaturing transforming growth factor-.beta. protein to provide an active fusion protein.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5800811
Application Number:US1995000470837
Date Filed:06/06/1995
Date Published:01/09/1998
Assignee:None


 
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