Isolated stromal cells and methods of using the same

• US 6,974,571 B2
• Filed: 03/28/1996
• Issued: 12/13/2005
• Est. Priority Date: 03/28/1995
• Status: Expired due to Fees

• Current Assignee: Thomas Jefferson University (Main Line Health System)
• Original Assignee: Thomas Jefferson University (Main Line Health System)
• Inventors: Prockop, Darwin J., Pereira, Ruth F., Laptev, Alexey, O'Hara, Michael D., Kulkosky, Joseph, Phinney, Donald, Leeper, Dennis B.
• Primary Examiner(s): Nguyen, Dave Trong
• Application Number:
  US08/913,918
• Publication Number:
  US 20030059412A1
• Time in Patent Office: 3,547 Days
• Field of Search: 424/93.21, 424/423, 424/425, 435/320.1, 435/69.1, 435/455, 435/325, 435/91.4
• US Class Current: 424/93.21
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 48/0025 wherein the non-active part clearly interacts with the delivered nucleic acid
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C12N 2502/1394 Bone marrow stromal cells; whole marrow
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Methods of treating patients who are suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect are disclosed. The methods comprising the step of intravenous administration of stromal cells isolated from normal syngeneic individuals or intravenous administration of stromal cells isolated from the patient subsequent to correction of the genetic defect in the isolated cells. Implant devices comprising a container that has at least one membrane surface and stromal cells isolated from bone marrow that comprise a gene construct are disclosed. The gene construct in the stromal cells comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells. Methods of treating individuals with diseases, disorders or conditions which can be treated with a beneficial protein, including diseases, disorders or conditions characterized by gene defects are disclosed. The methods comprise introducing into such individuals, stromal cells that are administered in a manner that physically isolates them from the recipient'"'"'s immune system and that comprise a gene construct that comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells.

Claims

1. An implantable container containing an isolated bone marrow stromal cell which comprises a first expressible gene construct encoding a protein, and a second expressible gene construct encoding a cytotoxic protein, which cytotoxic protein induces selective cell death in the presence of a drug specific for said cytotoxic protein, wherein said first and second expressible gene is under the control of a different promoter, further wherein the container physically isolates the stromal cell from immune cells of an animal when the container is implanted in the animal, and wherein the container has pores for permitting diffusion between the interior and the exterior of the container.

2. The container of claim 1, wherein the first gene construct encodes a secreted protein.

3. The container of claim 1, wherein the container is a microencapsulate stromal cell.

4. The container of claim 1, wherein the container is a biocompatible matrix having the stromal cells incorporated therein.

5. The container of claim 1, wherein the container comprises a membrane having pores which have a diameter not greater than about 0.3 micrometers.

6. The container of claim 5, wherein the container comprises a membrane having pores which have a diameter not greater than about 0.25 micrometers.

7. The container of claim 5, wherein the container comprises a membrane having pores which have a diameter not greater than about 0.1 micrometers.

8. The container of claim 1, wherein the stromal cell comprises a third expressible gene construct encoding a protein.

9. The container of claim 8, wherein the third expressible gene construct encodes an antibiotic resistance protein.

10. The container of claim 8, wherein the first expressible gene construct and the third expressible gene construct are the same gene construct.

11. The container of claim 1, wherein the stromal cell is a human stromal cell.

12. The container of claim 1, wherein the stromal cell is obtained from bone marrow.

13. The container of claim 1, containing at least 10⁴ of the stromal cells.

14. The container of claim 1, containing from 10⁴ to 10¹¹ of the stromal cells.

15. A method of providing a protein to an animal, the method comprising implanting within the animal a container containing an isolated marrow stromal cell which comprises a first expressible gene construct encoding a protein, and a second expressible gene construct encoding a cytotoxic protein, which cytotoxic protein induces selective cell death in the presence of a drug specific for said cytotoxic protein, wherein said first and second expressible gene is under the control of a different promoter, further wherein the container physically isolates the stromal cell from immune cells of the animal, and wherein the container has pores for permitting diffusion between the interior and the exterior of the container.

16. The method of claim 15, wherein said protein is a secreted protein.