Seeding parenchymal cells into compression resistant porous scaffold after vascularizing in vivo

• US 6,309,635 B1
• Filed: 11/28/1994
• Issued: 10/30/2001
• Est. Priority Date: 11/20/1986
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation, Massachusetts Institute of Technology
• Original Assignee: Children's Medical Center Corporation, Massachusetts Institute of Technology
• Inventors: Langer, Robert S., Vacanti, Joseph P., Ingber, Donald E.
• Primary Examiner(s): Naff, David M.
• Application Number:
  US08/345,217
• Time in Patent Office: 2,528 Days
• Field of Search: 435/177, 435/180, 435/240.21, 435/240.23, 435/240.243, 435/395, 435/396, 435/398, 435/402, 524/24, 524/421, 436/823, 424/422, 424/423, 424/456, 424/19, 424/93.7, 424/486, 423/11, 423/12, 423/13
• US Class Current: 424/93.7
• CPC Class Codes: A61F 2/022 Artificial gland structures using bioreactors hemodialysis A61M1/16; hemofiltration A61M1/34; other extracorporeal blood treatment A61M1/36
• 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 2430/06 for cartilage reconstruction, e.g. meniscus
• 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 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: C12N 2501/18 Liver cell growth factor (LCGF, Gly-His-Lys)
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• 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
• CPC Class Codes: C12N 5/0671 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

A method is provided whereby cells having a desired function are seeded on and into biocompatible, biodegradable or non-degradable porous polymer scaffolding matrix, previously implanted in a patient and infiltrated with blood vessels and connective tissue, to produce a functional organ equivalent. The resulting organoid is a chimera formed of parenchymal elements of the donated tissue and vascular and matrix elements of the host. The matrix should be compression resistant and a non-toxic, porous template for vascular ingrowth. The pore size, usually between approximately 100 and 300 microns, should allow vascular and connective tissue ingrowth throughout approximately 10 to 90% of the matrix, and the injection of cells such as hepatocytes without damage to the cells or patient. The introduced cells attach to the connective tissue and are fed by the blood vessels. Immediately prior to matrix implantation portacaval shunts can be created to provide trophic stimulatory factors to the implanted matrix to enhance replication and function.

Claims

1. A method enhancing in vivo survival of parenchymal cells in an implanted scaffold comprising:

a. implanting into the body of a patient a porous three-dimensional scaffold composed of a biocompatible polymer and having generally interconnected pores of between approximately 100 and 300 microns in diameter throughout the scaffold,

wherein the pores of the scaffold provide sufficient surface area to the scaffold to permit attachment of an amount of the cells effective to produce functional vascularized organ tissue in vivo,

wherein the scaffold is resistant to compression within the patient, to thereby maintain the pore size of the scaffold between approximately 100 and 300 microns, and

wherein the structure of the scaffold allows the introduction of cells into the vascularized scaffold without damage to the cells or patient;

b. maintaining the scaffold in the patient until the scaffold is between 10% and 90% vascularized and infiltrated with viable connective tissue; and

c. introducing viable parenchymal cells into the vascularized scaffold, wherein survival of the parenchymal cells in the vascularized scaffold is enhanced relative to survival of parenchymal cells in an unvascularized scaffold.

3. The method of claim 1 or 2 wherein the cells are introduced into the scaffold by means of a catheter.

4. The method of claim 1 or 2 wherein the scaffold is implanted in a tissue which is selected from the group consisting of the mesentery, subcutaneous tissue, subfascia, and supraperitoneal tissue.

5. The method of claim 1 or 2 further comprising performing a portacaval shunt on the patient.

6. The method of claim 1 or 2 wherein the scaffold is formed from a material selected from the group consisting of polyanhydride, polyorthoester, polyglycolic acid, polylactic acid, copolymers and blends thereof and collagen.

7. The method of claim 1 or 2 further comprising providing in the material forming the scaffold compounds selected from the group consisting of growth factors, compounds stimulating angiogenesis, and immunomodulators.

8. The method of claim 1 or 2 wherein the scaffold contains distribution channels for introduction of the cells.

9. The method of claim 1 or 2 further comprising providing with the scaffold means for introduction of the cells.

10. The method of claim 1 or 2 wherein the cells are hepatocytes.

11. The method of claim 1 or 2 further comprising selecting the cells from the group consisting of parathyroid cells, thyroid cells, cells of the adrenal-hypothalamic-pituitary axis, nerve cells, bone-forming cells, cells forming smooth muscle and cells forming skeletal muscle.

12. The method of claim 1 or 2, wherein the rate of survival of parenchymal cells is at least 40% after 24 hours.

13. The method of claim 1 or 2, wherein the rate of survival of parenchymal cells is in the range of 60-70% after 24 hours.

14. The method of claim 1 or 2, wherein the rate of survival of parenchymal cells is at least 25% after one week.

2. A method for enhancing in vivo survival of parenchymal cells in an implanted scaffold comprising:

a. implanting into the body of a patient a porous three-dimensional scaffold composed of a biocompatible polymer having a sponge or foam structure and having generally interconnected pores of between approximately 100 and 300 microns in diameter throughout the scaffold,

wherein the pores of the scaffold provide sufficient surface area to the scaffold to permit attachment of an amount of the cells effective to produce functional vascularized organ tissue in vivo,

wherein the scaffold is resistant to compression within the patient, to thereby maintain the pore size of the scaffold between approximately 100 and 300 microns, and

wherein the structure of the scaffold allows the introduction of cells into the vascularized scaffold without damage to the cells or patient;

b. maintaining the scaffold in the patient until the scaffold is between 10% and 90% vascularized and infiltrated with viable connective tissue; and

c. introducing viable parenchymal cells into the vascularized scaffold, wherein survival of the parenchytmal cells in the vascularized scaffold is enhanced relative to survival of parenchytmal cells in an unvascularized scaffold.

15. A method for producing a functional vascularized organ tissue in vivo comprising:

a. implanting into a patient a porous three-dimensional scaffold composed of a biocompatible polymer and having generally interconnected pores of between approximately 100 and 300 microns in diameter throughout the scaffold,

wherein the pores of the scaffold provide sufficient surface area to the scaffold to permit attachment of an amount of the cells effective to produce functional vascularized organ tissue in vivo, and

wherein the scaffold is resistant to compression within the patient, thereby maintaining the pore size of the scaffold to between approximately 100 and 300 microns, and the structure of the scaffold allows the introduction of cells into the vascularized scaffold without damage to the cells or patient;

b. maintaining the scaffold in the patient until the scaffold is between 10% and 90% vascularized and infiltrated with viable connective tissue; and

c. introducing viable hepatocytes into the vascularized scaffold.

16. A method for enhancing in vivo survival of hepatocytes in an implanted scaffold comprising:

a. implanting into the body of a patient a porous three-dimensional scaffold composed of a biocompatible polymer having a sponge or foam structure and having generally interconnected pores of between approximately 100 and 300 microns in diameter throughout the scaffold,

wherein the pores of the scaffold provide sufficient surface area to the scaffold to permit attachment of an amount of the cells effective to produce functional vascularized organ tissue in vivo,

wherein the scaffold is resistant to compression within the patient, to thereby maintain the pore size of the scaffold between approximately 100 and 300 microns, and

wherein the structure of the scaffold allows the introduction of cells into the vascularized scaffold without damage to the cells or patient;

b. maintaining the scaffold in the patient until the scaffold is between 10% and 90% vascularized and infiltrated with viable connective tissue; and

c. introducing viable hepatocytes into the vascularized scaffold, wherein survival of the hepatocytes in the vascularized scaffold is enhanced relative to survival of hepatocytes in an unvascularized scaffold.