Granted Patent B2

US 7,560,276 · App. 10/877,009 · Granted Jul 14, 2009

Abstract

Claims (19)

1. A method of stimulating tissue infiltration in a soft tissue defect in a patient in need thereof, said method comprising

(a) contacting mammalian umbilical cord tissue that is substantially free of blood with a composition comprising dispase and at least one of hyaluronidase and collagenase, thereby yielding isolated cells;

(b) optionally expanding said isolated cells in culture;

(c) placing said isolated or expanded, isolated cells onto a scaffold, thereby yielding a scaffold comprising cells; and then

(d) administering to said soft tissue defect of said patient said scaffold comprising cells;

wherein said isolated or expanded, isolated cells are capable of self-renewal and differentiation;

wherein said isolated or expanded, isolated cells do not express CD117; and

wherein said isolated or expanded, isolated cells exhibit increased expression relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an iliac crest bone marrow cell of an endogenous gene encoding oxidized low density lipoprotein receptor 1, an endogenous gene encoding interleukin 8, and an endogenous gene encoding reticulon 1; and

wherein said scaffold comprising cells stimulates tissue infiltration into said soft tissue defect.

2. The method of claim 1 wherein said isolated cells or said expanded, isolated cells require L-valine for growth and are capable of growth in about 5% to about 20% oxygen.

3. The method of claim 1 wherein said isolated cells or said expanded, isolated cells comprise the following characteristics:

potential for at least about 40 doublings in culture;

attachment and expansion on a coated or uncoated tissue culture vessel, wherein said coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyornithine, vitronectin, or fibronectin;

production of vimentin and alpha-smooth muscle actin; and

production of CD10, CD13, CD44, CD73, and CD90.

4. The method of claim 1 wherein the isolated cells or the expanded, isolated cells express each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A,B,C.

5. The method of claim 1 wherein the isolated cells or the expanded, isolated cells do not express any of CD31, CD34, CD45, CD117, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.

6. The method of claim 1 wherein the tissue comprises blood vessels.

7. The method of claim 1 wherein said step of administering comprises implanting, injecting, or engrafting said scaffold.

Assignments (8)