Methods of generating tissue using devitalized, acellular scaffold matrices derived from micro-organs

• US 7,297,540 B2
• Filed: 01/15/2002
• Issued: 11/20/2007
• Est. Priority Date: 01/15/2002
• Status: Active Grant

• Current Assignee: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. (Hebrew University of Jerusalem)
• Original Assignee: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. (Hebrew University of Jerusalem)
• Inventors: Mitrani, Eduardo N.
• Primary Examiner(s): Priebe; Scott D.
• Assistant Examiner(s): BERTOGLIO, VALARIE E
• Application Number:
  US10/045,018
• Publication Number:
  US 20030148510A1
• Time in Patent Office: 2,135 Days
• Field of Search: 435/1.1
• US Class Current: 435/395
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0688 Cells from the lungs or the respiratory tract

Abstract

Methods of generating, and isolating adult stem cells and utilizing such cells and/or embryonic stem cells in generating tissue of a specific function and micro-architecture are provided.

Claims

1. A method of generating an artificial micro-organ comprising:

(a) providing devitalized, acellular, tissue-derived three dimensional scaffold, said acellular three dimensional scaffold being of dimensions selected such that when populated with cells, said cells positionable deepest within said scaffold are at least about 100 micrometers and not more than about 225 micrometers away from said cells positioned at a nearest surface exposed to a source of gas and nutrients formed on said scaffold; and

(b) seeding said acellular three dimensional scaffold with stem cells, progenitor cells or differentiated cells, wherein said differentiated cells are of the same tissue from which the scaffold was generated, and

(c) providing conditions for cell growth and proliferation.

2. The method of claim 1, wherein said stem cells are adult stem cells.

3. The method of claim 1, wherein said stem cells are embryonic stem cells.

4. The method of claim 1, wherein said cells seeded on said acellular three dimensional scaffold are a mixed population of cells including stem cells, progenitor cells and differentiated cells, wherein said differentiated cells are of the same tissue from which the scaffold was generated.

5. The method of claim 1, wherein said cells seeded on said acellular three dimensional scaffold are genetically transformed to express at least one exogenous polypeptide.

6. The method of claim 1, wherein said stem cells are genetically transformed to express at least one exogenous polypeptide.

7. The method of claim 1, further comprising the step of generating said acellular three dimensional scaffold from a tissue-derived micro-organ.

8. The method of claim 7, wherein said step of generating is effected by subjecting said tissue-derived micro-organ to conditions selected suitable for removing cells and not acellular matrix from said micro-organ.

9. The method of claim 7, wherein said micro-organ is derived from an adult tissue of a type selected from the group consisting of skin, lung, kidney, liver and bone marrow.

10. The method of claim 7, wherein said progenitor cells are progenitor cells derived from the same tissue source as said micro-organ.