Platform for the differentiation of cells

• US 6,638,765 B1
• Filed: 08/03/2001
• Issued: 10/28/2003
• Est. Priority Date: 02/04/1999
• Status: Expired due to Fees

• Current Assignee: McGill University
• Original Assignee: McGill University
• Inventors: Rosenberg, Lawrence
• Primary Examiner(s): Fredman, Jeffrey
• Assistant Examiner(s): ANGELL, JON E
• Application Number:
  US09/890,717
• Time in Patent Office: 816 Days
• Field of Search: 435/366, 435/1.3, 435/380, 424/93.1
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/33 Insulin
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 2506/22 from pancreatic cells
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The present invention relates to an in vitro method for islet cell expansion, which comprises the steps of: a) preparing dedifferentiated cells derived from cells in or associated with post-natal islets of Langerhans; b) expanding the dedifferentiated cells; and c) inducing islet cell differentiation the expanded cells of step b) to become insulin-producing cells.

Claims

1. An in vitro method for islet cell expansion, which comprises the steps of:

a) inducing cystic formation in cells in or associated with post-natal islets of Langerhans to obtain a duct-like structure;

b) expanding cells of said duct-like structure; and

c) inducing islet cell differentiation of said expanded cells of said duct-like structure to become insulin-producing cells;

wherein step a) and step b) are concurrently effected using a solid matrix, basement membrane constituents, basal feeding medium and appropriate growth factors to permit the development, maintenance and expansion of a duct-like structure cell population with at least bipotentiality;

wherein step c) is effected by removing cells from said matrix and resuspended in a basal liquid medium containing soluble matrix proteins and growth factors selected from the group consisting of fibronectin, IGF-1, IGF-2, insulin, and NGF.

2. The method of claim 1, wherein said basal liquid medium is CMRL 1066 supplemented with 10% fetal calf serum.

3. The method of claim 1, wherein said basal liquid medium further comprises glucose concentration of at least 11 mM.

4. The method of claim 3, wherein said basal liquid medium further comprises inhibitors of known intracellular signaling pathways of apoptosis and/or specific inhibitor of p38.