Biomatrix Composition and Methods of Biomatrix Seeding

Abstract

Apparatus and methods are described for generating autologous tissue grafts, the apparatus including a point of care SVF isolation unit that includes a tissue digestion chamber in fluid communication with a lipid separating chamber, whereby SVF cells are isolated without centrifugation; and a cell seeding chamber in fluid communication with the SVF isolation unit, said cell seeding chamber adapted to support a cell scaffold. Methods and materials for cell seeding, including through the provision of micro rough scaffold surfaces, are also provided.

We claim: 
     
         1 . A cell-seeded tissue graft comprising a reparative cell preparation seeded onto a porous scaffold, wherein the tissue graft is prepared by:
 isolating a fresh stromal vascular fraction (SVF) from an adipose tissue of a patient by a process including enzymatically digesting the adipose tissue;   applying the fresh SVF cells to the porous scaffold without subjecting the fresh SVF cells to plastic adherence; and   rinsing the porous scaffold to eliminate cells that are unbound by the porous scaffold, thereby generating a tissue graft comprising a reparative cell preparation in less than about 4 hours from removal of the adipose tissue from the patient.   
     
     
         2 . The cell-seeded tissue graft of  claim 1 , wherein the porous scaffold is characterized by a micro-rough cell attachment surface that has surface irregularities at a periodicity of 1-20 μm. 
     
     
         3 . The cell-seeded tissue graft of  claim 1 , wherein the cell-seeded tissue graft is generated at a point-of-care and is implanted into the patient without culturing the cell-seeded tissue graft. 
     
     
         4 . The cell-seeded tissue graft of  claim 1 , wherein the cell-seeded tissue graft is cultured to expand populations of cells seeded on the graft prior to implanting into the patient. 
     
     
         5 . The cell-seeded tissue graft of  claim 1 , wherein the SVF cells are pushed into contact with the porous scaffold by pressure. 
     
     
         6 . The cell-seeded tissue graft of  claim 1 , wherein the SVF cells are pushed into contact with the porous scaffold by a partial vacuum. 
     
     
         7 . The cell-seeded tissue graft of  claim 1 , wherein the SVF cells are incubated with an inductive media before, during or after being applied to the porous scaffold. 
     
     
         8 . The cell-seeded tissue graft of  claim 7 , wherein the inductive media is adapted for generation of one or more of adipocytes, chondrocytes, endothelial cells, hepatocytes, myoblasts, neurons, and osteoblasts. 
     
     
         9 . The cell-seeded tissue graft of  claim 1 , wherein the porous scaffold is comprised of a biocompatible or a biodegradable material. 
     
     
         10 . The cell-seeded tissue graft of  claim 9 , wherein the biocompatible material is selected from the group consisting of: polytetrafluoroethylene, woven polyester, spun silicone, open foam silicone encased in microporous expanded PTFE, stainless steel, polypropylene, polyurethane, polycarbonate, nickel titanium shape memory alloys and cobalt-chromium-nickel alloys, and combinations thereof. 
     
     
         11 . The cell-seeded tissue graft of  claim 10 , wherein the biodegradable material is selected from the group consisting of: silk fibroin-chitosan, acellular dermal matrices, collagen, polyglactin, hyaluronic acid, and resorbable silica gel matrix. 
     
     
         12 . The cell-seeded tissue graft of  claim 2 , wherein the surface irregularities of the porous scaffold are created by treatment of at least one cell attachment surface of the scaffold by one or more of embossing, blasting, plasma etching, by controlling polymerization or drying processes, by heat application, by chemical etching, and by coating or printing. 
     
     
         13 . The cell-seeded tissue graft of  claim 2 , wherein at least one surface of the porous scaffold is characterized by a spongy texture formed by subjecting the nascent scaffold material to a partial vacuum during polymerization or drying. 
     
     
         14 . The cell-seeded tissue graft of  claim 1 , wherein the cell-seeded tissue graft is adapted and configured to treat one or more of: wound healing, burns, bone fractures, cosmetic defects, cartilage damage, tendon damage, ulcers, fistulas, hernias, retinal degeneration, treatment of ischemic disease, nerve injury, aneurysms, bladder wall repair, intestinal injury, and vascular vessel repair. 
     
     
         15 . The cell-seeded tissue graft of  claim 1 , wherein an adherence agent selected to promote adherence of desired cell types is applied to the porous scaffold prior to cell-seeding. 
     
     
         16 . The cell-seeded tissue graft of  claim 15 , wherein the adherence agent is selected from the group consisting of: autologous plasma or serum and components thereof, cold insoluble globulin, carboxymethyl dextran, iron dextran, and hyaluronic acid and polymers thereof. 
     
     
         17 . A cell-seeded tissue graft comprising a reparative cell preparation seeded onto a porous scaffold, wherein the tissue graft is prepared by:
 isolating a fresh stromal vascular fraction (SVF) from extraction fluid of a lipoaspirate from a patient;   applying the fresh SVF cells to the porous scaffold without subjecting the fresh SVF cells to plastic adherence; and   rinsing the porous scaffold to eliminate cells that are unbound by the porous scaffold, thereby generating a tissue graft comprising a reparative cell preparation in less than about 4 hours from removal of the adipose tissue from the patient.