In vitro platform for screening agents inducing islet cell neogenesis

• US 20070009882A1
• Filed: 05/27/2004
• Published: 01/11/2007
• Est. Priority Date: 05/27/2003
• Status: Abandoned Application

• Current Assignee: McGill University
• Original Assignee: McGill University
• Inventors: Rosenberg, Lawrence
• Application Number:
  US10/558,191
• Publication Number:
  US 20070009882A1
• Time in Patent Office: Days
• US Class Current: 435/4
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2506/22 from pancreatic cells
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: G01N 2333/4701 Details
• CPC Class Codes: G01N 2400/36 beta-D-Fructofuranans, e.g. levan, insulin
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5017 for testing neoplastic activity
• CPC Class Codes: G01N 33/5023 on expression patterns
• CPC Class Codes: G01N 33/5026 on cell morphology
• CPC Class Codes: G01N 33/507 Pancreatic cells

Abstract

The present invention relates to an in vitro method for screening agents inducing islet cell neogenesis or duct-to-islet cell transdifferentiation, which comprises the steps of: a) expanding in vitro cells of a duct-like structure obtained by inducing cystic formation in cells in or associated with post-natal islets of Langerhans; b) treating said expanded cells of said duct-like structure with an agent screened; and c) determining potency of said agent of inducing islet cell differentiation of said duct-like structure in becoming insulin-producing cells.

Claims

1. An in vitro method for screening agents inducing islet cell neogenesis or duct-to-islet cell transdifferentiation, which comprises the steps of:

a) expanding in vitro cells of a duct-like structure obtained by inducing cystic formation in cells in or associated with post-natal islets of Langerhans;

b) treating said expanded cells of said duct-like structure with an agent being screened; and

c) determining potency of said agent of inducing islet cell differentiation of said duct-like structure in becoming insulin-producing cells.

2. The method of claim 1, wherein step a) and step b) are concurrently effected using a solid matrix, basal feeding medium and appropriate growth factors to permit the development, maintenance and expansion of a dedifferentiated cell population with at least bipotentiality.

3. The method of claim 2, wherein said solid matrix is 3-D collagen type-1 gel matrix, said basal liquid medium is DMEM/F12 medium supplemented with EGF and cholera toxin.

4. The method of claim 1, wherein said cells are human cells.

5. A kit for carrying out the method of claim 1, which comprises:

a) a solid matrix for 3-D culture of cells;

b) a culture medium supplemented.

8. An islet cell culture, which comprises insulin-producing islet cells in a suitable culture medium, wherein said islet cells are characterized.

6. The kit of claim 8, wherein sais solid matrix is 3-D collagen type-1 gel matrix and said medium is DMEM/F12 medium supplemented with EGF and cholera toxin.

7. The kit of claim 8, which further comprises duct-like structure cells or islet cells to be transformed into duct-like structure cells.

9. The islet cell culture of claim 8, wherein said characterization is genetic, immunologic or genomic.

10. The islet cell culture of claim 9, wherein said characterization is effected using a DNA microarray analysis.

11. An in vitro method for evaluating biological effects of agents on islet cells, which comprises the steps of:

a) treating the islet cell culture of any one of claims 8 to 10 with an agent being evaluated for a time sufficient for a biological effect to be occurring; and

b) determining biological effect of said agent on islet cells by monitoring changes in insulin production compared to, a standard curve obtained with a control islet cell culture.

12. The method of claim 11, wherein said agent is selected from the group consisting of immunosuppressive agents, growth factors and anti-apoptotic agents.