Methods for in vitro expansion and transdifferentiation of human pancreatic acinar cells into insulin-producing cells

• US 20060275900A1
• Filed: 04/26/2006
• Published: 12/07/2006
• Est. Priority Date: 05/28/2002
• Status: Abandoned Application

• Current Assignee: Becton, Dickinson & Co, Novocell, Inc. (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Becton, Dickinson & Co, Novocell, Inc. (Vertex Pharmaceuticals Incorporated)
• Inventors: Scharp, David William, Presnell, Sharon C., Heidaran, Mohammad, Coutts, Margaret, McIntyre, Catherine Anne, Haaland, Perry, Latta, Paul Presley
• Application Number:
  US11/410,954
• Publication Number:
  US 20060275900A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• 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 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/46 Amines, e.g. putrescine
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/315 : Prolactin
• CPC Class Codes: C12N 2501/335 : Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/34 : Calcitonin; Calcitonin-gene related peptide [CGRO]; Amylin
• CPC Class Codes: C12N 2501/345 : Gastrin; Cholecystokinins [CCK]
• CPC Class Codes: C12N 2501/35 : Vasoactive intestinal peptide [VIP]; Pituitary adenylate cyclase activating polypeptide [PACAP]
• CPC Class Codes: C12N 2501/37 : Parathyroid hormone [PTH]
• CPC Class Codes: C12N 2501/39 : Steroid hormones
• CPC Class Codes: C12N 2501/392 : Sexual steroids
• CPC Class Codes: C12N 2501/41 : Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/83 : Tachykinins, e.g. substance P
• CPC Class Codes: C12N 2501/85 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/22 : from pancreatic cells
• CPC Class Codes: C12N 5/0037 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

This invention relates, e.g., to a method for expanding mammalian acinar cells, comprising culturing the cells in a cell culture system comprising a cell culture medium and a cell attachment surface, under conditions wherein the acinar cells undergo a 3-4 fold expansion together with transdifferentiation into a modified cell phenotype (IP cells) showing characteristics of acinar cells and liver cells. The invention also relates to a method for transforming these IP cells to insulin-producing cells in vitro, comprising culturing the cells in a novel, defined medium. Also disclosed are suitable culture media for performing these methods, isolated cells having the phenotype of IP cells and/or produced by these methods, and kits for performing the methods.

Claims

1-10. -10. (canceled)

11. An isolated insulin-producing cell generated by a method comprising the step of culturing an IP cell in a cell culture medium comprising an effective amount of at least one differentiation promoting factor selected from the group consisting of C-Natriuretic Peptide (CNP), Calcitonin Gene Related Peptide, Cholera Toxin B Subunit, Dexamethasone, Gastrin-Releasing Peptide, Laminin, Met-Enkephalin, PDGFAA+PDGFBB, Sonic Hedgehog, and Substance P, such that the IP cell is transformed into an insulin-producing cell.

12. An insulin-producing cell, prepared by differentiating a mammalian acinar cell in vitro, wherein said insulin-producing cell has an expression profile after 16 days ex vivo as shown in Table 6.

13-20. -20. (canceled)