Localized delivery of factors enhancing survival of transplanted cells

• US 6,281,015 B1
• Filed: 12/16/1994
• Issued: 08/28/2001
• Est. Priority Date: 12/16/1994
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation, Massachusetts Institute of Technology
• Inventors: Langer, Robert S., Vacanti, Joseph P., Mooney, David J.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US08/358,235
• Time in Patent Office: 2,447 Days
• Field of Search: 424/93.7, 424/457, 424/462, 435/325, 435/395, 435/405, 514/3
• US Class Current: 435/395
• CPC Class Codes: A61K 35/407 Liver; Hepatocytes
• CPC Class Codes: A61K 38/1808 Epidermal growth factor [EGF] urogastrone
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2531/00 Microcarriers
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0075 using microcarriers
• CPC Class Codes: C12N 5/067 Hepatocytes

Abstract

Growth factors and/or angiogenic factors are administered in combination with dissociated cells to be transplanted, preferably in microspheres with the cells on or in a polymeric matrix, to enhance survival and proliferation of the transplanted cells. Examples demonstrate that epidermal growth factor (EGF) was incorporated into microspheres fabricated from a copolymer of lactic and glycolic acid using a double emulsion technique, the incorporated EGF was steadily released over one month in vitro, and it remained biologically active, as determined by its ability to stimulate DNA synthesis, division, and long-term survival of cultured hepatocytes. EGF-containing microspheres were mixed with a suspension of hepatocytes, seeded onto porous sponges, and implanted into the mesentery of two groups of Lewis rats, to demonstrate efficacy in vivo. Two weeks after implantation in PCS animals, devices which included EGF-containing microspheres showed a two-fold increase in the number of engrafted hepatocytes, as compared to implants which received blank microspheres.

Claims

1. A method for delivering a bioactive factor to a transplantation site, said method comprising introducing a microsphere comprising a bioactive factor to a hepatocyte-seeded biodegradable, biocrodible polymer matrix prior to transplantation, wherein said bioactive factor is exposed to portal circulation factors, and wherein said bioactive factor is selected from the group consisting of a growth factor, a factor inhibiting ingrowth of fibrous tissue, and a factor inhibiting cancerous growth, whereby the controlled release of said bioactive factor over time modulates the microenvironment of the transplanted hepatocyte to improve engraftment of the hepatocyte transplanted at the site.

2. The method of claim 1, wherein the factors are selected from the group consisting of heparin binding growth factor, transforming growth factor alpha or beta, alpha fibroblastic growth factor, epidermal growth factor, vascular endothelium growth factor, insulin, glucagon, and estrogen.

3. The method of claim 1, wherein said polymer matrix is a hydrogel polymer solution.

4. The method of claim 3, wherein said cell is seeded in said polymer matrix by suspending it in said hydrogel polymer solution.

5. The method of claim 1, wherein said polymer matrix is a fibrous structure.

6. The method of claim 5, wherein said fibrous structure has interstitial spacing between 100 and 300 microns.

7. The method of claim 1, wherein said transplanted cell is exposed to portal circulation factors by perfoming a portal caval shunt at the time of or prior to implantation of said cells.

8. The method of claim 1, wherein said microsphere comprises a polymer.

9. The method of claim 8, wherein said polymer is comprised of poly(lactide), poly(glycolic acid), poly(lactide-co-glycolide), poly(caprolactone), polycarbonate, polyamide, polyanhydride, polyamino acid, polyolthoester, polyacetal, polycyanoacrylate, polyurethane, polyacrylate, ethylene-vinyl acetate, acyl substituted cellulose acetate, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonated polyolefin, or polyethylene oxide.