In vitro growth of functional islets of Langerhans

• US 5,834,308 A
• Filed: 04/28/1994
• Issued: 11/10/1998
• Est. Priority Date: 04/28/1994
• Status: Expired due to Fees

• Current Assignee: University of Florida (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Peck, Ammon B., Cornelius, Janet G.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US08/234,071
• Time in Patent Office: 1,657 Days
• Field of Search: 435/240.1, 435/240.2, 435/240.21, 435/240.3, 435/240.31, 435/70.1, 435/70.3, 435/325, 435/354, 435/366, 435/371, 435/41
• US Class Current: 435/325
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• 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: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The subject invention concerns new methods which make it possible, for the first time, to grow functional islet cells in in vitro cultures. The ability to grow these cells opens up important new avenues for research and therapy relating to diabetes.

Claims

1. A method for the production of islet cells comprising culturing mammalian pancreatic stem cells in a nutrient medium supplemented with serum from the same mammalian species, growing the stem cells for at least three weeks allowing for the establishment of mature stromal cells and initiating the differentiation of said stromal cells into islet cells by adding serum from the same mammalian species.

2. The method according to claim 1, wherein the islet cells are human islet cells and the serum is human serum.

3. The method according to claim 1, wherein the islet cells are mouse islet cells and the serum is mouse serum.

4. The method according to claim 1, wherein differentiation of cultured stem cells is initiated at about 4 to 5 weeks of culture growth by addition of serum.

5. The method according to claim 1 wherein said nutrient medium comprises a high amino acid nutrient medium.

6. The method according to claim 1 further comprising, after the step of initiating differentiation by added serum, refeeding with said serum at about one-week intervals.

7. A method for producing insulin comprising culturing mammalian pancreatic stem cells in a nutrient medium supplemented with serum from the same mammalian species, growing the stem cells for at least three weeks allowing for the establishment of mature stromal cells, initiating the differentiation of said stromal cells into islet cells by adding serum from the same mammalian species and recovering the insulin.

8. The method according to claim 7, wherein said insulin is human insulin.

9. The method according to claim 7, wherein said insulin is mouse insulin.

10. The method according to claim 7, wherein differentiation is initiated at about 4 to 5 weeks of culture growth by refeeding with medium comprising normal serum.