Neomorphogenesis of urological structures in vivo from cell culture

• US 5,851,833 A
• Filed: 08/07/1996
• Issued: 12/22/1998
• Est. Priority Date: 10/24/1991
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Atala, Anthony
• Primary Examiner(s): Lankford, Jr., Leon B.
• Assistant Examiner(s): TATE, CHRISTOPHER ROBIN
• Application Number:
  US08/692,243
• Time in Patent Office: 867 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/240.241-241.243, 435/325, 435/378, 435/380, 435/395-402, 424/422, 424/423, 424/426, 424/486
• US Class Current: 435/378
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61L 2430/22 for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3882 Hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Methods and artificial matrices for the growth and implantation of urological structures and surfaces are disclosed in which urothelial cells are grown in culture on biodegradable, biocompatible, fibrous matrices formed of polymers, such as polyglycolic acid, polylactic acid, or other polymers which degrade over time. The cells can be cultured in vitro until an adequate cell volume and density has developed for the cells to survive and proliferate in vivo. Alternatively, when adequate cell numbers for implantation are available, the cells can be attached to the matrix and implanted directly, without proliferation in vitro. The implants approximate the desired urological structure to be replaced or repaired, such as the kidney, urether, bladder, urethra, and the like. Implantation is followed by remodeling through cell growth and proliferation in vivo. In another aspect of the invention, techniques are disclosed for selectively extracting or harvesting urothelial cells either from excised urological tissue in vitro or from intact urological tissue in vivo by treating the tissue with a digestive enzyme, such as collagenase.

Claims

1. A method for isolating and culturing normal mammalian urothelial cells, the method comprising:

exposing urological tissue to an enzyme solution;

collecting normal mammalian urothelial cells which are dissociated by the solution; and

culturing the dissociated urothelial cells on a biocompatible, biodegradable polymeric matrix suitable for implantation at a site where urothelial tissue is to be repaired or replaced.

2. The method of claim 1 wherein the enzyme solution is a collagenase solution.

3. The method of claim 2 wherein the concentration of collagenase in the solution ranges from about 0.05 percent to about 0.75 percent.

4. The method of claim 1 wherein the step of exposing the tissue further comprises excising a tissue biopsy, isolating the mucosa from the biopsy and placing the mucosa in the enzyme solution.

5. The method of claim 1 wherein the step of exposing the tissue further comprises irrigating the interior of a bladder in vivo with the enzyme solution.

6. The method of claim 1 wherein the step of collecting urothelial cells further comprises centrifuging a solution containing the dissociated cells.

7. The method of claim 1 wherein the method comprises:

providing the biocompatible, biodegradable polymeric matrix in a shape suitable for repair of kidney or bladder and attaching urothelial cells to the matrix.

8. The method of claim 7 wherein the method comprises providing the biocompatible, biodegradable polymeric matrix in a shape suitable for reconstruction or repair of a tubular structure.