Postpartum cells derived from umbilical cord tissue, and methods of making, culturing, and using the same

• US 20060171930A1
• Filed: 12/08/2005
• Published: 08/03/2006
• Est. Priority Date: 12/21/2004
• Status: Abandoned Application

• Current Assignee: Advanced Technologies and Regenerative Medicine, LLC
• Original Assignee: Advanced Technologies and Regenerative Medicine, LLC
• Inventors: Gosiewska, Anna, Seyda, Agnieszka
• Application Number:
  US11/297,156
• Publication Number:
  US 20060171930A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3641 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0654 Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0691 Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels

Abstract

Cells derived from human umbilical cords are disclosed along with methods for their therapeutic use. Isolation techniques, culture methods and detailed characterization of the cells with respect to their cell surface markers, gene expression, and their secretion of trophic factors are described.

Claims

1. A method for forming a cell sheet, which comprises coating a support with a polymer or copolymer which has a lower or upper critical solution temperature within the range of 0°

C. to 80°

C., culturing cells attached to said surface thereof at a higher temperature than said lower critical solution temperature or at a lower temperature than said upper critical solution temperature on said coated surface, and detaching and collecting said cell sheet from said coated surface by changing the temperature to a lower temperature than said lower critical solution temperature or to a higher temperature than said upper critical solution temperature;

wherein said cells are isolated umbilicus-derived cells comprising cells derived from mammalian umbilical cord tissue substantially free of blood, said cells capable of self-renewal and expansion and having the potential to differentiate into cells of other phenotypes.

2. The method of claim 1 wherein said polymer or copolymer comprises the reaction products of monomers selected from the group consisting of acrylamide, methacrylamide, N-ethyl acrylamide, N-n-propyl acrylamide, N-n-propyl methacrylamide, N-isopropyl acrylamide, N-isopropyl methacrylamide, N-cyclopropyl acrylamide, N-cyclopropyl methacrylamide, N-ethoxyethyl acrylamide, N-ethoxyethyl methacrylamide, N-tetrahydrofurfuryl acrylamide, N-tetrahydrofurfuryl methacrylamide, N,N-dimethyl(meth)acrylamide, N,N-ethylmethyl acrylamide, N,N-diethyl acrylamide), 1-(1-oxo-2-propenyl)-pyrrolidine, 1-(1-oxo-2-propenyl)-piperidine, 4-(1-oxo-2-propenyl)-morpholine, 1-(1-oxo-2-methyl-2-propenyl)-pyrrolidine, 1-(1-oxo-2-methyl-2-propenyl)-piperidine, 4-(1-oxo-2-methyl-2-propenyl)-morpholine, and methyl vinyl ether.

3. The method of claim 2 wherein said polymer comprises the reaction product of N-isopropyl acrylamide.

4. The method of claim 1 wherein said support is comprised of a material selected from the group consisting of polystyrene, poly(methyl methacrylate)), polypropylene, polyethylene, vinyl polymers, ceramics, metals, glass and modified glass.

5. The method of claim 1 wherein said isolated umbilicus-derived cells are free of maternal cells.

6. The method of claim 1 wherein said cells are comprised of a co-culture comprised of isolated umbilicus-derived cells comprising cells derived from mammalian umbilical cord tissue substantially free of blood and another mammalian cell of any phenotype.

7. The method of claim 6 wherein said another mammalian cell comprise a human cell line.

8. A method for forming sheets of isolated umbilicus-derived cells, which comprises coating a support with a polymer which has a lower critical solution temperature of less than about 30°

C., culturing said cells at 37°

C. to confluence, lowering the temperature to 20°

C., and collecting said sheets.

9. The method of claim 8 wherein said isolated umbilicus-derived cells comprise cells derived from mammalian umbilical cord tissue substantially free of blood, said cells capable of self-renewal and expansion and having the potential to differentiate into cells of other phenotypes.

10. The method of claim 8 wherein said polymer comprises the reaction product of N-isopropyl acrylamide.

11. The method of claim 8 wherein said support is comprised of a material selected from the group consisting of polystyrene, poly(methyl methacrylate)), ceramics, metals, glass and modified glass.

12. The method of claim 8 wherein said isolated umbilicus-derived cells are free of maternal cells.

13. A method for therapeutically treating mammalian tissue, said method comprising the steps of

I. providing a cell sheet, the cell sheet formed by a method which comprises coating a support with a polymer or copolymer which has a lower or upper critical solution temperature within the range of 0°

C. to 80°

C., culturing cells attached to said surface thereof at a higher temperature than said lower critical solution temperature or at a lower temperature than said upper critical solution temperature on said coated surface, and detaching and collecting said cell sheet from said coated surface by changing the temperature to a lower temperature than said lower critical solution temperature or to a higher temperature than said upper critical solution temperature, wherein said cells are isolated umbilicus-derived cells comprising cells derived from mammalian umbilical cord tissue substantially free of blood, said cells capable of self-renewal and expansion and having the potential to differentiate into cells of other phenotypes; and

,

II. transplanting the cell sheet to mammalian tissue.

14. The method of claim 13 wherein said polymer or copolymer comprises the reaction products of monomers selected from the group consisting of acrylamide, methacrylamide, N-ethyl acrylamide, N-n-propyl acrylamide, N-n-propyl methacrylamide, N-isopropyl acrylamide, N-isopropyl methacrylamide, N-cyclopropyl acrylamide, N-cyclopropyl methacrylamide, N-ethoxyethyl acrylamide, N-ethoxyethyl methacrylamide, N-tetrahydrofurfuryl acrylamide, N-tetrahydrofurfuryl methacrylamide, N,N-dimethyl(meth)acrylamide, N,N-ethylmethyl acrylamide, N,N-diethyl acrylamide), 1-(1-oxo-2-propenyl)-pyrrolidine, 1-(1-oxo-2-propenyl)-piperidine, 4-(1-oxo-2-propenyl)-morpholine, 1-(1-oxo-2-methyl-2-propenyl)-pyrrolidine, 1-(1-oxo-2-methyl-2-propenyl)-piperidine, 4-(1-oxo-2-methyl-2-propenyl)-morpholine, and methyl vinyl ether.