Endothelial cell procurement and deposition kit

• US 5,035,708 A
• Filed: 05/24/1989
• Issued: 07/30/1991
• Est. Priority Date: 06/06/1985
• Status: Expired due to Term

• Current Assignee: Thomas Jefferson University (Main Line Health System)
• Original Assignee: Becton, Thomas Jefferson University (Main Line Health System)
• Inventors: Cutshall, Tony A., Williams, Stuart K., Alchas, Paul G., Brooks, Christopher J., Gabel, Jonathan B., Mulhauser, Paul J., Augello, Frank A., Jarrell, Bruce E., Rose, Deborah G., DiPisa, Joseph A. Jr., Prais, Wes
• Primary Examiner(s): Shay, Randy Citrin
• Application Number:
  US07/356,431
• Time in Patent Office: 797 Days
• Field of Search: 623/1, 623/12, 623/15, 600/36, 604/4, 604/5, 604/19, 604/35, 604/48, 604/248, 604/319, 604/902, 128/752, 435/1, 435/240.21, 435/290, 30/210, 210/258, 210/217
• US Class Current: 623/1.45
• CPC Class Codes: A61F 2/06 Blood vessels
• 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 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/36 containing ingredients of undetermined constitution or reaction products thereof , e.g. transplant tissue, natural bone, extracellular matrix isolated natural compounds, e.g. collagen A61L27/24
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: A61M 2202/0014 removed from the body
• CPC Class Codes: A61M 2202/08 Lipoids
• CPC Class Codes: C08L 89/06 derived from leather or skin , e.g. gelatin
• CPC Class Codes: C12M 45/09 by enzymatic treatment
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: Y10S 604/902 Suction wands

Abstract

The invention is an endothelial cell procurement and deposition kit for collecting fat from a patient, processing said fat to produce an endothelial cell deposition product, and depositing said product on the surface of a graft, all under sterile conditions established and maintained within the components of said kit comprised of: fat collection means for collecting subcutaneous fat from a patient; digestion means connectable to said fat collection means to maintain sterility during reception of said fat and for retaining said fat under sterile conditions during rinsing and digestion to produce a digested product; endothelial cell isolation means connectable to said digestion means for maintaining sterile conditions during reception of said digested product and for separating and isolating microvessel endothelial cells from said digested product to produce an endothelial cell product; cell deposition means connectable to said isolation means for maintaining sterile conditions during reception of said endothelial cell product and for depositing said cells on the surface of a graft to be implanted in a patient and facilitating implantation of said endothelial graft into a patient.

Claims

1. An endothelial cell procurement and deposition kit for collecting fat from a patient, processing said fat to produce an endothelial cell deposition product, and depositing said product on the surface of a graft, all under sterile conditions established and maintained within the components of said kit, comprising:

a fat collection means for collecting fat from a patient;

digestion means connectable to said fat collection means to maintain sterility during reception of said fat and for retaining said fat under sterile conditions during rinsing and digestion to produce a digested product;

endothelial cell isolation means connectable to said digestion means for maintaining sterile conditions during reception of said digested product and for separating and isolating microvessel endothelial cells from said digested product to produce an endothelial cell product; and

cell deposition means connectable to said isolation means for maintaining sterile conditions during reception of said endothelial cell product and for depositing said cells on the surface of a graft to be implanted in a patient and facilitating implantation of said endothelialized graft into a patient.

2. The kit of claim 1 wherein said fat collection means further comprises fat receiving means connectable to said fat collection means for receiving said collected fat and for storing said fat under sterile conditions until needed.

3. The kit of claim 2 wherein said fat receiving means further comprises:

a chamber having a top and a bottom;

stopper means substantially in a sealing engagement with the chamber capable of being slidably raised and lowered with said chamber;

inlet tube means connectable to fat collection means extending through said stopper means into said chamber;

vacuum inlet tube means extending through said stopper means into said chamber;

plunger means connected to said stopper means for lowering and raising said stopper means;

diaphragm means affixed to said stopper means for sealing said chamber during fat transfer, said diaphragm means being deflected by said inlet tube means and said vacuum inlet tube means when said stopper means is in a raised position and closing off both said inlet means when the stopper means is in a lowered position thereby facilitating fat transfer from the chamber;

outlet means affixed to the bottom of the chamber including a valve means for regulating the transfer of fat from the chamber

4. The kit of claim 1 wherein said fat collection means comprises liposuction means.

5. The kit of claim 1 wherein said digestion means further comprises:

a chamber having a top and a bottom;

a first inlet port affixed at the top of the chamber connectable to the fat collection means for introducing collected fat into the digestion means chamber;

a second inlet port affixed at the top of the chamber for introducing rinsing solution into the digestion means chamber;

a third inlet port affixed at the top of the chamber for introducing digesting solution into the digestion means chamber;

a fourth inlet port affixed at the top of the chamber connectable to a vacuum source;

a fifth inlet port affixed at the top of the chamber for introducing filtered ambient air into the digestion means chamber;

a tube centrally disposed within the chamber parallel to the longitudinal axis of the chamber said tube having a top and a bottom, and connected at its top end to said fifth inlet port and connected at its bottom end to an agitating means, through which tube filtered ambient air can be introduced into said chamber;

agitating means connected to the said bottom end of said central tube;

filtering means connectable under the agitating means for retaining undigested debris;

monitoring means connectable at the bottom of the chamber for monitoring the reaction within the chamber;

a first outlet port affixed at the bottom of the chamber for removing rinse solution and digestion debris;

a second outlet port affixed at the bottom of the chamber connectable to the endothelial cell isolation means for transferring said digested product to the endothelial cell isolation means.

6. The kit of claim 5 wherein said monitoring means further comprises:

a protective sheath containing a temperature probe connectable to an external heat controller whereby the reaction temperature within the digestion means is controlled.

7. The kit of claim 5 wherein said agitating means further comprises:

a plurality of elongated members affixed to the centrally disposed tube whereby filtered air introduced through the tube is distributed for uniform agitation and a frictional surface to facilitate fat digestion is provided.

8. The kit of claim 1 wherein said endothelial cell isolation means comprises:

a first chamber connectable to the digestion means for receiving digested product having a top and a bottom;

a first inlet port at the top of said first chamber connectable to the digestion means;

a second chamber connected to the bottom of the first chamber to isolate the endothelial cell product;

a second inlet port affixed to said second chamber having a two-position valve allowing communication between said first and second chamber when in a first position and communication between said second chamber and said inlet port to isolate endothelial cell product when in a second position;

an outlet port affixed to said second chamber having a second two-position valve allowing communication with the portion of said second chamber above said second two-position valve and the portion of said second chamber below said second two position valve to isolate isolated endothelial cell product from packed red blood cells when in a second, closed position;

whereby when said isolation means containing said digested product is centrifuged, the endothelial cells are isolated from the digested product within said second chamber when both of said two position valves are in a second, closed position.

9. The kit of claim 1 wherein said cell deposition means comprises:

an inner process tube for supporting the graft during deposition of endothelial cell product onto the graft under sterile conditions; and

an outer process tube for housing the inner process tube, and maintaining sterility during the cell deposition process.

10. The cell deposition means of claim 9 wherein said outer process tube further comprises:

a tube body;

an endcap connectable to the tube body having an inlet means connectable to the endothelial cell isolation means and a communicating outlet means connectable to the inlet means of the inner process tube;

an endcap connectable to the tube body having communicating inlet and outlet means wherein said inlet means is connectable to the outlet means of the inner process tube.

11. The cell deposition means of claim 10 wherein said outer process tube further comprises:

rotary fittings connectable to both end caps whereby the device may be rotated about its central axis.

12. The cell deposition means of claim 9 wherein said inner process tube further comprises:

a tube body;

a tunneler assembly connectable to the tube body and to the outer process tube at the end which is connectable to the cell isolation means;

a vent cap connectable to the tube body having an outlet means connectable to the outer process tube end cap;

whereby cell product introduced under pressure flows through the graft housed in the tunneler assembly and deposits cell product onto the graft.

13. The inner process tube of claim 12 wherein the tunneler assembly housing the graft comprises:

a tunneler for supporting the graft;

a tunneler tip connectable to the tunneler for facilitating proper graft placement within the patient;

a handling cap connectable to the tunneler, and to the graft housed within the tunneler, and to the inner process tube body to facilitate handling of the tunneler assembly.

14. The inner process tube of claim 13 wherein said tunneler further comprises:

a tube having a plurality of holes disposed along its axis whereby endothelial cell product flows through the graft housed within the tunneler and then out through said holes depositing endothelial cell product onto the graft.

15. A method for sterilely procuring and depositing endothelial cells onto a graft to be implanted in a patient, said method comprising the steps of:

providing a kit for collecting fat from a patient, processing said fat to produce an endothelial cell deposition product, and depositing said product on the surface of a graft, all under sterile conditions established and maintained within the components of said kit including a fat collection means for collecting fat from a patient, a digestion means for producing a digested product, an endothelial cell means for producing an endothelial cell product and a cell deposition means for depositing said endothelial cell product on said graft;

collecting fat from a patient into said sterile fat collection means;

sterilely transferring said collected fat into said sterile digestion means;

rinsing and digesting said fat within said digestion means to produce a digested product;

sterilely transferring said digested product into said sterile isolation means;

isolating endothelial cells from said digested product within said isolation means to produce an endothelial cell product;

sterilely transferring said endothelial cell product into said sterile cell deposition means; and

sterilely depositing said cell product onto the surface of the graft to be implanted into a patient.

16. The method of claim 15 wherein the step of isolating endothelial cells from digested product further comprises centrifuging said isolation means to isolate said endothelial cell product.

17. The method of claim 15 wherein the step of depositing endothelial cell product onto a graft further comprises:

creating negative pressure external to the graft wherein said graft is porous; and

flowing endothelial cell product into the graft lumen thereby depositing cell product onto the graft surface.

18. The method of claim 17 wherein the method of depositing endothelial cell product onto the surface of the graft further comprises rotating the graft during the cell deposition process.