Method of differentiating mammalian progenitor cells into insulin producing pancreatic islet cells

• US 8,877,497 B2
• Filed: 05/21/2009
• Issued: 11/04/2014
• Est. Priority Date: 05/22/2008
• Status: Active Grant

• Current Assignee: Vesta Therapeutics, Inc.
• Original Assignee: Vesta Therapeutics, Inc.
• Inventors: Ruiz, Joseph Charles, Hoynowski, Steven Michael
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US12/470,247
• Publication Number:
  US 20100136688A1
• Time in Patent Office: 1,993 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2500/30 Organic components
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/065 Modulators of histone acetylation
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/14 from hepatocytes
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: G01N 33/5014 for testing toxicity
• CPC Class Codes: G01N 33/5038 involving detection of metabolites per se
• CPC Class Codes: G01N 33/507 Pancreatic cells

Abstract

The invention relates to methods for differentiating progenitor cells into insulin producing pancreatic islet cells and compositions and methods for using such cells.

Claims

1. A method for differentiating hepatic progenitor cells into insulin-producing pancreatic cells, comprising:

(a) incubating hepatic progenitor cells expressing albumin and CK19 in serum-free pre-induction media comprising sodium butyrate; and

, subsequently,

(b) incubating the hepatic progenitor cells in induction media comprising knock-out replacement serum;

for a time sufficient to differentiate the hepatic progenitor cells into insulin-producing pancreatic cells.

2. The method according to claim 1 in which the hepatic progenitor cells are human hepatic progenitor cells.

3. The method according to claim 1 in which the induction media comprises about 2% knock-out replacement serum.

7. The method according to claim 1 in which the serum-free pre-induction media comprises 1-5 mM sodium butyrate.

4. A method for differentiating hepatic progenitor cells into insulin-producing pancreatic cells, comprising:

(a) incubating hepatic progenitor cells expressing albumin and CK19 in serum-free pre-induction media comprising a histone deacetylase inhibitor (HDACI) selected from the group consisting of phenyl butyrate, retinoic acid, valproic acid, APHA Compound 8, Apicidin, (−

)-Depudecin, Scriptaid, Sirtinol, and Trichostatin; and

, subsequently,

(b) incubating the hepatic progenitor cells in induction media comprising knock-out replacement serum;

for a time sufficient to differentiate the hepatic progenitor cells into insulin-producing pancreatic cells.

5. The method according to claim 4 in which the HDACI is selected from the group consisting of retinoic acid, valproic acid, APHA Compound 8, Apicidin, (−

)-Depudecin, Scriptaid, Sirtinol, and Trichostatin.

6. The method according to claim 4 in which the hepatic progenitor cells are human hepatic progenitor cells.