Differentiation of non-embryonic stem cells to cells having a pancreatic phenotype

• US 8,409,859 B2
• Filed: 10/16/2006
• Issued: 04/02/2013
• Est. Priority Date: 10/14/2005
• Status: Active Grant

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Verfaillie, Catherine M., Heremans, Yves Pierre, Barajas Velez, Miguel Angel
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US12/089,868
• Publication Number:
  US 20090092586A1
• Time in Patent Office: 2,360 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/19 Growth and differentiation factors [GDF]
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2506/03 from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The invention provides methods for differentiating non-embryonic multipotent stem cells along the pancreatic lineage. The present invention further provides non-embryonic multipotent stem cells and progeny derived therefrom to provide pancreatic cells to a subject.

Claims

1. An in vitro method comprising contacting non-embryonic stem, non-germ, non-embryonic germ cells that can differentiate into at least two of ectodermal, endodermal and mesodermal cell types with agents consisting essentially of Activin A and an agent that inhibits sonic hedgehog (SHH) activity to yield cells having increased expression of Pdx-1.

2. The method of claim 1 further comprising contacting the non-embryonic stem, non-germ, non-embryonic germ cells with factors consisting essentially of BMP4.

3. The method of claim 1 further comprising contacting the cells having increased Pdx-1 expression with factors consisting essentially of EGF or HGF to yield cells having increased expression of Ngn3.

4. An in vitro method to differentiate non-embryonic stem, non-germ, non-embryonic germ cells that can differentiate into at least two of ectodermal, endodermal and mesodermal cell types comprising the steps of:

a) contacting the non-embryonic stem, non-germ and non-embryonic germ cells with factors consisting essentially of Activin A and an agent that inhibits sonic hedgehog activity, to produce cells with increased expression of Pdx-1;

b) contacting the cells obtained from step a) with factors consisting essentially of EGF or HGF, to produce cells expressing Ngn3; and

c) contacting the cells obtained from step b) with factors consisting essentially of one or both of nicotinamide or exendin4, to produce cells expressing insulin.

5. The method of claim 4 wherein step a), step b) or both further comprise contacting the cells with factors consisting essentially of BMP4.

6. The method of claim 4 wherein step d) further comprises contacting the cells with factors consisting essentially of one or both of GDF11 or betacellulin.

7. A composition comprising (1) factors consisting essentially of Activin A and an agent that inhibits sonic hedgehog and (2) non-embryonic stem, non-germ, non-embryonic germ cells that differentiate into at least two of ectodermal, endodermal and mesodermal cell types.

8. The composition of claim 7 further comprising factors consisting essentially of BMP4.

9. The composition of claim 7, wherein the composition further comprises cell culture medium or a pharmaceutically acceptable carrier.

10. The composition of claim 7 wherein the sonic hedgehog inhibitor is cyclopamine or an anti-SHH antibody.

11. The composition of claim 8, wherein the composition further comprises cell culture medium or a pharmaceutically acceptable carrier.