Implantable prosthetic vascular device having an adherent cell monolayer produced under shear stress

• US 5,843,781 A
• Filed: 01/04/1996
• Issued: 12/01/1998
• Est. Priority Date: 04/28/1993
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Ott, Mark J., Ballermann, Barbara J.
• Primary Examiner(s): Naff, David M.
• Application Number:
  US08/583,490
• Time in Patent Office: 1,062 Days
• Field of Search: 435/240.242, 435/240.243, 435/395, 435/396, 435/400, 435/402, 435/180, 435/363, 435/364, 435/373, 424/422, 424/424, 424/93.3, 424/93.7
• US Class Current: 435/400
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: C12M 25/10 Hollow fibers or tubes hollow fiber modules in general B01D63/02
• CPC Class Codes: C12N 5/0686 Kidney cells
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells

Abstract

A device having a surface coated with a firmly adherent cell monolayer is produced by culturing adherent cells in the presence of the surface in vitro under conditions of continuous shear stress of from 0.4 dyne/cm² to 33 dyne/cm² produced by the force of circulating fluid medium in contact with the cells. The surface may be contained by an implantable device, or a culture or fermentation vessel. Preferably, an endothelial cell monolayer is produced on a surface of a prosthetic vascular device made of polypropylene. In a hollow fiber cartridge device, endothelial cells are grown under shear stress on the inner surface of the lumen of a hollow fiber and perivascular cells are grown on the outer surface of the fiber. Growing cells under continual stress more closely approximates the in vivo environment where blood passes over the endothelium in a blood vessel, and produces a cell monolayer closely resembling naturally occurring firmly adherent cell layers found in vivo in the lining of blood vessels. The endothelial cells may be transfected with a gene and the surface of the device may be pre-coated with extracellular matrix protein.

Claims

1. A method for producing an implantable prosthetic vascular device comprising providing a hollow fiber cartridge device containing at least one hollow fiber having an inner lumen surface inside the fiber and an outer lumen surface outside the fiber, seeding endothelial cells on the inner lumen surface and perivascular cells on the outer lumen surface, culturing the endothelial cells under a continuous shear stress of from 0.4 dyne/cm² to 33 dyne/cm² to produce an adherent endothelial cell monolayer on the inner lumen surface and culturing the perivascular cells to produce a layer of perivascular cells on the outer lumen surface.

2. The method of claim 1, wherein the perivascular cells are selected from the group consisting of vascular smooth muscle cells, mesangial cells, pericytes, fibroblasts and epithelial cells.

3. The method of claim 1, wherein the hollow fiber is made of polypropylene material.

4. The method of claim 1, wherein the endothelial cells are stably transfected with at least one gene.

5. The method of claim 4, wherein the gene encodes a protein which reduces thrombogenicity.

6. The method of claim 5, wherein the gene is selected from the group consisting of t-PA and PLA₂.

7. The method of claim 4, wherein the gene encodes a vasodilating mediator.

8. The method of claim 4, wherein the gene encodes an angiogenic factor.

9. The method of claim 1, wherein the inner lumen surface is pre-coated with an extracellular matrix protein.

10. The method of claim 9, wherein the extracellular matrix protein is selected from the group consisting of fibronectin, laminin, and collagen.

11. An implantable prosthetic vascular device produced by the method of claim 1.