METHOD FOR DIFFERENTIATING EMBRYONIC STEM CELLS INTO CELLS EXPRESSING AQP-1

• US 20110070280A1
• Filed: 07/21/2008
• Published: 03/24/2011
• Est. Priority Date: 07/19/2007
• Status: Active Grant

• Current Assignee: Annegret Schumacher, Gunter Maubach, Jackie Y. Ying, Karl M. Schumacher, Karthikeyan Narayanan
• Original Assignee: Annegret Schumacher, Gunter Maubach, Jackie Y. Ying, Karl M. Schumacher, Karthikeyan Narayanan
• Inventors: Narayanan, Karthikeyan, Schumacher, Karl M., Schumacher, Annegret, Maubach, Gunter, Ying, Jackie Y.
• Granted Patent: US 8,481,316 B2
• Time in Patent Office: Days
• US Class Current: 424/422
• CPC Class Codes: A61F 2/06 Blood vessels
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0686 Kidney cells

Abstract

The present invention relates to methods of differentiating a human embryonic stem (ES) cell into a cell, specifically a renal epithelial cell, expressing AQP-I. The methods disclosed comprise culturing human ES cells in a renal specific medium in the presence of an extracellular matrix molecule. The cells produced according to said method can be used to treat renal related disorders such as renal failure, nephrosis, Bright'"'"'s disease and glomerulitis.

Claims

1. A method of differentiating a human embryonic stem (hES) cell into a cell expressing AQP-1, the method comprising:

culturing an undifferentiated hES cell in a renal-specific culture medium in the presence of an extracellular matrix (ECM) molecule, under conditions sufficient to induce differentiation of the hES cell to a cell expressing AQP-1.

2. The method according to claim 1 wherein the renal-specific culture medium comprises renal epithelial basal medium or renal epithelial growth medium.

3. The method according to claim 1 wherein the ECM molecule comprises at least one of fibronectin, laminin, collagen IV and Matrigel matrix.

4. The method according to claim 1 wherein the renal-specific culture medium comprises an epithelial growth factor.

5. The method according to claim 1 further comprising adding bone morphogenetic protein 2 to the renal-specific culture medium before or during said culturing.

6. The method according to claim 1 further comprising adding bone morphogenetic protein 2 to the renal-specific culture medium at a concentration of from about 2.5 to about 10 ng/ml before or during said culturing, wherein said culturing comprises growing the cell for 7-10 days, the renal-specific culture medium comprises renal epithelial growth medium and the ECM molecule comprises Matrigel matrix, and wherein once differentiated, the cell expresses AQP-1 and at least one of CK-18, β

-catenin, CD-326 and CD-133.

8. The population of cells according to claim 7, when prepared by a method of claim 1.

7. A population of cells differentiated directly from a population of human embryonic stem cells in a renal-specific culture medium and in the presence of an extracellular matrix molecule, wherein about 2% to about 50% of the cells express AQP-1.

9. A bioartificial tubule assist device comprising a hollow core fibre having an interior lumen, the interior lumen coated with an extracellular matrix (ECM) molecule and a population of cells of claim 7.

10. A method of preparing a bioartificial tubule assist device, the method comprising providing a bioartificial tubule assist device comprising a hollow core fibre having an interior lumen and seeding the interior lumen with a population of cells of claim 7, wherein the interior lumen is coated with an extracellular matrix molecule prior to seeding with the population of cells.

11. The method according to claim 10 wherein the interior lumen is coated with an extracellular matrix molecule comprising at least one of fibronectin, laminin, collagen IV and Matrigel matrix.

12. The method according to claim 10, wherein the population of cells is cultured to a confluent monolayer coating the interior lumen.

13. A method of treating a renal related disorder in a subject in need thereof, the method comprising implanting an effective amount of a population of cells of claim 7 in the subject at a site where cells expressing AQP-1 are required.

14. The method according to claim 13 wherein the population of cells is provided within a bioartificial tubule assist device of claim 9.

15. A method of treating a renal related disorder in a subject comprising externally connecting a bioartificial tubule assist device of claim 9 to a subject in need thereof.

16. The method according to claim 13 wherein the renal related disorder comprises renal failure, nephropathy, diabetic nephropathy, nephrosis, Bright'"'"'s disease, renal insufficiency, glomerulitis, glomerulosclerosis or nephritis.