Methods for reprogramming cells and uses thereof
AssigneeGenesis Technologies Limited
InventorsJan-Eric Ahlfors, Rouwayda El-Ayoubi
Life of this patent
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Abstract
Described herein are reprogrammed cells, and methods for cell dedifferentiation, transformation and eukaryotic cell reprogramming. Also described are cells, cell lines, and tissues that can be transplanted in a patient after steps of in vitro dedifferentiation and in vitro reprogramming. In particular embodiments the cells are Stem-Like Cells (SLCs), including Neural Stem-Like Cells (NSLCs), Cardiac Stem-Like Cells (CSLC), Hematopoietic Stem-Like Cells (HSLC), Pancreatic Progenitor-Like Cells, and Mesendoderm-like Cells. Also described are methods for generating these cells from human somatic cells and other types of cells. Also provided are compositions and methods of using of the cells so generated in human therapy and in other areas.
Claims9
An in vitro method of transforming a cell of a first type to a hematopoietic multipotent cell, comprising:
- i) providing a cell of a first type, wherein the cell of the first type is a somatic cell that is multipotent, unipotent or terminally differentiated, but not a hematopoietic multipotent cell;
- ii) introducing into said cell of a first type reprogramming agent(s) or polynucleotides encoding said reprogramming agents) selected from the group consisting of:
- (1) Brachyury (T), Caudal Type Homeobox 4 (CDX4), Homeobox B4 (HOXB4), GATA Binding Factor 1 (GATA1), GATA Binding Factor 2 (GATA2), Kruppel-like Factor 1 (KLF1), Left-right determination factor 1 (LEFTY1), Eomesodermin (FOMES), Delta-like protein 1 precursor (DLL1), GATA Binding Factor 4 (GATA4), GATA Binding Factor 6 (GATA6), Heart and Neural Crest Derivatives Expressed-1 (HAND1), Insulin gene enhancer protein (ISL1), and NK2 Homeobox 5 (NKX2.5);
- (2) Eomesodermin;
- (3) CDX4;
- (4) KLF1;
- (5) Eomesodermin combined with Gata1, Gata2, Klf1, Cdx4, Tal1, Bmi1, Fli1, or HoxB4;
- (6) Cdx4 combined with Gata1, Gata2 or Klf1;
- (7) Gata1 combined with Fli1, Cdx4, HoxB4 or Eomesodermin;
- and
- (8) HoxB4 combined with Tal1, Fli1, Gata1, or Eomesodermin;
- iii) placing the cell of the first type in a hematopoietic cell culture medium and maintaining expression of the reprogramming agent(s) or polynucleotides encoding said reprogramming agent(s);
- and
- iv) maintaining the cell in hematopoietic cell culture medium for a sufficient period of time to allow a stable expression of a plurality of secondary genes whose expression is characteristic of phenotypical and/or functional properties of the hematopoietic multipotent cell, wherein at least one of said secondary genes is not characteristic of phenotypical and functional properties of an embryonic stem cell, whereby at the end of said period of time the cell of the first type has been transformed into the hematopoietic multipotent cell.
An in vitro method of obtaining a hematopoietic multipotent cell, comprising:
- i) providing a cell of a first type, wherein the cell of the first type is a somatic cell that is multipotent, unipotent or terminally differentiated;
- ii) contacting chromatin and/or DNA of the cell of a first type with a histone acetylator, an inhibitor of histone deacetylation, a DNA demethylator, and/or an inhibitor of DNA methylation;
- iii) introducing into said cell of a first type reprogramming agent(s) or polynucleotides encoding said reprogramming agent(s) selected from the group consisting of:
- (1) Brachyury (T), Caudal Type Homeobox 4 (CDX4), Homeobox B4 (HOXB4), GATA Binding Factor 1 (GATA1), GATA Binding Factor 2 (GATA2), Kruppel-like Factor 1 (KLF1), Left-right determination factor 1 (LEFTY1), Eomesodermin (FOMES), Delta-like protein 1 precursor (DLL1), GATA Binding Factor 4 (GATA4), GATA Binding Factor 6 (GATA6), Heart and Neural Crest Derivatives Expressed-1 (HAND1), Insulin gene enhancer protein (ISL1), and NK2 Homeobox 5 (NKX2.5);
- (2) Eomesodermin;
- (3) CDX4;
- (4) KLF1;
- (5) Eomesodermin combined with Gata1, Gata2, Klf1, Cdx4, Tal1, Bmi1, Fli1, or HoxB4;
- (6) Cdx4 combined with Gata1, Gata2 or Klf1;
- (7) Gata1 combined with Fli1, Cdx4, HoxB4 or Eomesodermin;
- and
- (8) HoxB4 combined with Tal1, Fli1, Gata1, or Eomesodermin;
- and
- iv) culturing the cell of a first type in a hematopoietic cell culture medium and maintaining stable expression of the reprogramming agent(s), wherein the reprogramming agent(s) is capable of driving directly or indirectly transformation of the cell of the first type into the hematopoietic multipotent cell whereby a hematopoietic multipotent cell is obtained.
Dependencies are inferred from the claim text.