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US 10,179,151 B2Granted
Methods of transdifferentiation and methods of use thereof
AssigneesOrgenesis Inc., Tel HaShomer Medical Research Infrastructure and Services Ltd.
InventorSarah Ferber
Life of this patent
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Abstract
Disclosed herein is a method for manufacturing a population of human insulin producing cells from non-pancreatic β-cells, wherein the resulting insulin producing cells have increased insulin content, or increased glucose regulated secretion of insulin, or a combination of both.
Claims26
A method of manufacturing a population of human insulin producing cells, the method comprising the steps of:
- (a) obtaining adult human liver tissue;
- (b) processing said liver tissue to recover primary adult human liver cells;
- (c) propagating and expanding said primary adult human liver cells to a predetermined number of cells;
- (d) transdifferentiating said expanded cells, wherein said transdifferentiating comprises:
- (1) infecting said expanded cells with at least one expression vector, said infecting occurring at a first time period,
- (i) wherein said at least one expression vector comprises an adenoviral vector comprising a nucleic acid encoding a PDX-1 polypeptide and an expression vector comprising an adenoviral vector comprising a nucleic acid encoding a second pancreatic transcription factor polypeptide selected from NeuroD1 and Pax4, wherein said infecting with said adenoviral vectors occurs at the same time,
- or
- (ii) wherein said at least one expression vector comprises an adenoviral vector comprising a nucleic acid encoding a PDX-1 polypeptide and a second pancreatic transcription factor polypeptide selected from NeuroD1 and Pax4;
- and
- (2) infecting said expanded infected cells of (1) with an adenoviral vector comprising a nucleic acid encoding a MafA polypeptide, said infecting occurring at a second time period, wherein said second time period is after said first time period;
- and
- (e) harvesting said transdifferentiated expanded cells;
- thereby manufacturing said population of human insulin producing cells having an increased insulin content, or increased glucose regulated insulin secretion, or increased glucose regulated C-peptide secretion, or any combination thereof, compared with control non-transdifferentiated liver cells.
10 more elements
A population of human insulin producing cells manufactured by a method comprising the steps of:
- (a) obtaining adult human liver tissue;
- (b) processing said liver tissue to recover primary adult human liver cells;
- (c) propagating and expanding said primary adult human liver cells to a predetermined number of cells;
- (d) transdifferentiating said expanded cells, wherein said transdifferentiating comprises:
- (1) infecting said expanded cells with at least one expression vector, said infecting occurring at a first time period,
- (i) wherein said at least one expression vector comprises an adenoviral vector comprising a nucleic acid encoding a PDX-1 polypeptide and an expression vector comprising an adenoviral vector comprising a nucleic acid encoding a second pancreatic transcription factor polypeptide selected from NeuroD1 and Pax4, wherein said infecting with said adenoviral vectors occurs at the same time,
- or
- (ii) wherein said at least one expression vector comprises an adenoviral vector comprising a nucleic acid encoding a PDX-1 polypeptide and a second pancreatic transcription factor polypeptide selected from NeuroD1 and Pax4;
- and
- (2) infecting said expanded infected cells of (1) with an adenoviral vector comprising a nucleic acid encoding a MafA polypeptide, said infecting occurring at a second time period, wherein said second time period is after said first time period;
- and
- (e) harvesting said transdifferentiated expanded culture;
- wherein said population of human insulin producing cells have an increased insulin content or increased glucose stimulated insulin secretion, or increased glucose regulated C-peptide secretion, or any combination thereof, compared with control non-transdifferentiated liver cells.
10 more elements
Dependencies are inferred from the claim text.