US2005003537A1PendingUtilityA1

Composition and method for production of transformed cells

47
Priority: Feb 17, 1995Filed: Nov 24, 2003Published: Jan 6, 2005
Est. expiryFeb 17, 2015(expired)· nominal 20-yr term from priority
Inventors:Stephen F. BadylakJeffrey BonadioSherry Voytik
A61F 2210/0004A61K 48/0025A61K 35/38A61L 27/3629A61K 48/00A61F 2002/30062A61F 2002/30677A61F 2002/2835A61F 2/28A61L 27/38
47
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Claims

Abstract

A composition useful for the production of transformed eukaryotic cells is described. The composition comprises submucosal tissue and a nucleic acid sequence. The nucleic acid sequence is typically recombinant DNA including gene(s) encoding for one or more biofunctional proteins. The submucosal tissue component of the present composition comprises the tunica submucosa of vertebrate intestine delaminated from the tunica muscularis and at least the luminal portion of the tunica mucosa. Injection or implantation of the composition into a host induces the formation of transformed cells capable of expressing gene(s) encoded by the nucleic acid sequence.

Claims

exact text as granted — not AI-modified
1 . A method for inducing the production of eukaryotic cells containing exogenous nucleic acid sequences, said method comprising the step of contacting target cells with a transformation composition comprising submucosal tissue of a warm-blooded vertebrate and an exogenous nucleic acid sequence, under conditions conducive to the proliferation of said target cells.  
     
     
         2 . The method of  claim 1 , wherein the submucosal tissue is intestinal submucosa comprising the tunica submucosa delaminated from the tunica muscularis and at least the luminal portions of the tunica mucosa of warm-blooded vertebrate intestine.  
     
     
         3 . The method of  claim 1 , wherein the step of contacting cells is conducted in vivo and comprises the step of implanting the transformation composition into a host vertebrate species.  
     
     
         4 . The method of  claim 1 , wherein the transformation composition is in an injectable form and is injected into a host to contact host cells.  
     
     
         5 . The method of  claim 1 , wherein the transformation composition comprises a nucleic acid sequence and intestinal submucosal tissue consisting essentially of the tunica submucosa, the muscularis mucosa and stratum compactum of the intestine of a warm-blooded vertebrate.  
     
     
         6 . The method of  claim 1 , wherein the exogenous nucleic acid sequence is selected from the group consisting of an RNA sequence or a DNA sequence.  
     
     
         7 . The method of  claim 6 , wherein the exogenous nucleic acid sequence is an antisense nucleic acid sequence.  
     
     
         8 . The method of  claim 6 , wherein the exogenous nucleic acid sequence is circular.  
     
     
         9 . The method of  claim 6 , wherein the nucleic acid sequence comprises a gene operably linked to regulatory sequences for expressing the gene in the target cells.  
     
     
         10 . The method of  claim 6 , wherein the DNA sequence comprises a sequence encoding antisense RNA operably linked to regulatory sequences for expressing the encoded antisense RNA in a eukaryotic cell.  
     
     
         11 . The method of  claim 9 , wherein the transformation composition is implanted or injected in a vertebrate host to induce the production of host cells containing the DNA sequence, wherein said DNA sequence encodes a host-deficient cellular product.  
     
     
         12 . The method of  claim 10 , wherein the transformation composition is implanted or injected into a vertebrate host to induce the production of host cells containing the DNA sequence.  
     
     
         13 . The method of  claim 2 , wherein the intestinal submucosa comprises the tunica submucosa, the muscularis mucosa and the stratum compactum of the tunics mucosa of a segment of small intestine of a warm-blooded vertebrate, said tunica submucosa, muscularis mucosa and stratum compactum being delaminated from the tunica muscularis and the luminal portion of the tunica mucosa of said segment.  
     
     
         14 . A composition comprising submucosal tissue of a warm-blooded vertebrate and a nucleic acid.  
     
     
         15 . The composition of  claim 14 , wherein the submucosal tissue is intestinal submucosa comprising the tunica submucosa delaminated from the tunica muscularis and at least the luminal portions of the tunica mucosa of warm-blooded vertebrate intestine.  
     
     
         16 . The composition of  claim 15 , wherein said nucleic acid comprises a DNA sequence encoding a gene for a biofunctional protein operably linked to regulatory sequences for expressing the gene in a eukaryotic cell.  
     
     
         17 . The composition of  claim 15 , wherein said nucleic acid sequence is circular.  
     
     
         18 . The composition of  claim 17 , wherein said circular nucleic acid sequence is a plasmid.  
     
     
         19 . The composition of  claim 15 , wherein the intestinal submucosa consists essentially of the tunica submucosa and basilar tissue of the tunica mucosa of the intestine of a warm-blooded vertebrate.  
     
     
         20 . The composition of  claim 15  in injectable form.  
     
     
         21 . The composition of  claim 20 , wherein the intestinal submucosa is solubilized by partial hydrolysis.  
     
     
         22 . A method of preparing the composition of  claim 14 , said method comprising the steps of contacting the submucosal tissue with a solution of the nucleic acid sequence.  
     
     
         23 . The method of  claim 22 , further comprising the step of desiccating the submucosal tissue prior to contacting the tissue with the nucleic acid sequence solution.  
     
     
         24 . An in vivo transformation composition comprising a DNA sequence and intestinal submucosal tissue, said intestinal tissue comprising the tunica submucosa and the basilar tissue of the tunica mucosa of vertebrate small intestine, wherein the DNA sequence encodes at least one gene operably linked to regulatory sequences for expressing the gene in eukaryotic cells.  
     
     
         25 . The transformation composition of  claim 21 , wherein said nucleic acid sequence is circular.  
     
     
         26 . The transformation composition of  claim 25 , wherein said circular nucleic acid sequence is a plasmid.  
     
     
         27 . An injectable, non-immunogenic tissue graft comprising comminuted or solubilized submucosal tissue in combination with a DNA sequence encoding a gene for a biofunctional protein.  
     
     
         28 . A method for inducing the production of eukaryotic cells containing exogenous nucleic acid sequences, said method comprising the step of contacting target cells with a transformation composition comprising a vertebrate derived collagenous matrix and an exogenous nucleic acid sequence, under conditions conducive to the proliferation of said target cells.  
     
     
         29 . The transformation composition of  claim 28 , wherein said nucleic acid sequence is a plasmid.  
     
     
         30 . The method of  claim 28 , wherein the exogenous nucleic acid sequence is an antisense nucleic acid sequence.  
     
     
         31 . The method of  claim 28 , wherein the nucleic acid sequence comprises a gene operably linked to regulatory sequences for expressing the gene in the target cells.

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