Method for differentiating embryonic stem cells into cells expressing AQP-1

• US 8,481,316 B2
• Filed: 07/21/2008
• Issued: 07/09/2013
• Est. Priority Date: 07/19/2007
• Status: Expired due to Fees

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: Schumacher, Karl M., Ying, Jackie Y., Schumacher, Annegret, Narayanan, Karthikeyan, Maubach, Gunter
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US12/669,457
• Publication Number:
  US 20110070280A1
• Time in Patent Office: 1,814 Days
• Field of Search: 435/377, 435/366, 435/384
• US Class Current: 435/377
• 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, CD-326 and CD-133, the method comprising:

culturing an undifferentiated hES cell in a renal epithelial medium that comprises (i) serum and (ii) an epithelial growth factor 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, CD-326 and CD-133.

2. The method according to claim 1 wherein the renal epithelial 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 epithelial medium comprises renal epithelial growth medium and the epithelial growth factor and the serum are components of the renal epithelial growth medium.

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

6. The method according to claim 1 further comprising adding bone morphogenetic protein 2 to the renal epithelial 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 epithelial medium comprises renal epithelial growth medium and the ECM molecule comprises Matrigel matrix, and wherein once differentiated, the cell expresses AQP-1, CD-326 and CD-133 and at least one of CK-18 and β

-catenin.