Method for generating non-pluripotent progenitors of surrogate pancreatic cells

• US 9,474,772 B2
• Filed: 04/22/2015
• Issued: 10/25/2016
• Est. Priority Date: 06/26/2012
• Status: Active Grant

• Current Assignee: Seraxis, Inc. (Seraxis Pte Ltd.)
• Original Assignee: Seraxis, Inc. (Seraxis Pte Ltd.)
• Inventors: Rust, William L.
• Primary Examiner(s): Lankford, Blaine
• Application Number:
  US14/693,245
• Publication Number:
  US 20150231181A1
• Time in Patent Office: 552 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• 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 2500/05 Inorganic components
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/30 Organic components
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• 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 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2502/1329 Cardiomyocytes
• CPC Class Codes: C12N 2506/22 from pancreatic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

Fresh human pancreas tissue can be used as a source of cells whence to identify and select a non-stem cell population that is predisposed to be a source for surrogate pancreatic cells that can be used in treating insulin-dependent diabetes. The progenitors of these surrogate pancreatic cells have no reprogramming genes integrated into their genomes, differentiate to the pancreatic lineage pursuant to a protocol that employs only defined reagents, and are substantially unable to differentiate to the mesodermal lineage.

Claims

1. A method for generating a composition comprising non-pluripotent progenitors of surrogate pancreatic cells that are suitable for treating insulin-dependent diabetes, comprising:

a. harvesting human cells from viable human pancreatic tissue in minimal, defined culture conditions;

b. culturing the primary human cells for a period of days that is fewer than about 9 days;

then

c. delivering reprogramming genes into the primary human cells from (b) to reprogram the primary human cells to obtain reprogrammed cells such that no reprogramming genes are integrated into the genome of the reprogrammed cells, wherein the reprogrammed cells have a stem cell morphology;

d. firstly selecting among the reprogrammed cells obtained in (c) for an ability to proliferate, without losing said stem cell morphology, in minimal, defined culture conditions whereby proliferating reprogrammed cells are obtained; and

then

e. secondly selecting from among the proliferating reprogrammed cells for a cell population characterized by (i) an ability to survive and differentiate to a pancreatic lineage in the course of a protocol that employs only defined reagents, and (ii) a substantial inability to differentiate to a mesodermal lineage, whereby said non-pluripotent progenitors are obtained.