Granted Patent B2

US 12,624,342 · App. 18/188,421 · Granted May 12, 2026

Inventors: Jan-Eric Ahlfors (Laval, CA); Rouwayda El-Ayoubi (Laval, CA)

Assignee: Genesis Technologies Limited

C12N5/0662A61K35/30C12N5/0618C12N5/0619C12N5/0623C12N5/0647C12N5/0656C12N5/0657C12N5/0667C12N5/0668C12N5/0696A61K35/545C12N5/0676C12N2500/25C12N2501/06C12N2501/065C12N2501/105C12N2501/11C12N2501/115C12N2501/13C12N2501/155C12N2501/16C12N2501/395C12N2501/60C12N2501/602C12N2501/604C12N2501/727C12N2501/998C12N2506/094C12N2506/11C12N2506/1307C12N2506/1346C12N2506/1384C12N2510/00C12N2513/00

Abstract

Claims (21)

1 . A method of obtaining a cell having pluripotent-like or multipotent characteristics, comprising:

i) providing a cell of a first type, which is not a cell having pluripotent-like or multipotent characteristics;

ii) contacting the cell of a first type with an agent capable of remodeling the chromatin and/or DNA of the cell, wherein the agent capable of remodeling the chromatin and/or DNA is a histone acetylator, an inhibitor of histone deacetylation, a DNA demethylator, and/or an inhibitor of DNA methylation;

iii) introducing a nucleic acid encoding at least one pluripotent gene regulator in the cell of a first type, to achieve an expression level of the at least one pluripotent gene regulator that is capable of driving transformation of the cell of a first type into the cell having pluripotent-like or multipotent characteristics, wherein the at least one pluripotent gene regulator comprises Nanog, Rex1, Lin28, Tpt1, DPPA4, or a combination thereof; and

iv) placing or maintaining the cell in a differentiation medium and maintaining intracellular levels of the at least one pluripotent gene regulator for a sufficient period of time to allow a stable cell having pluripotent-like or multipotent characteristics to be obtained;

wherein the cell having pluripotent-like or multipotent characteristics so obtained does not exhibit teratoma formation in vivo.

2 . The method of claim 1 , wherein, in step (ii), the remodeling agent is methyl-CpG binding domain protein 2 (MBD2), Growth arrest and DNA-damage-inducible beta (Gadd45b), valproic acid or 5-azacytidine.

3 . The method of claim 1 , wherein, in step (ii), the remodeling agent is methyl-CpG binding domain protein 2 (MBD2).

4 . The method of claim 1 , wherein the cell having pluripotent-like or multipotent characteristics so obtained expresses one or more pluripotent-like cell marker selected from the group consisting of Oct4, Sox2, Nanog, SSEA-4, TRA1-60, TRA1-81 and AP.

5 . The method of claim 1 , wherein the cell having pluripotent-like or multipotent characteristics so obtained possesses all of the following characteristics:

(i) can self-renew for significantly longer than a somatic cell;

(ii) is not a cancerous cell;

(iii) is stable and not artificially maintained by forced gene expression and may be maintained in standard cell media;

(v) can differentiate to a unipotent or somatic cell; and

(vi) does not exhibit uncontrolled growth or tumor formation in vivo.

6 . The method of claim 1 , wherein a plurality of cells having pluripotent-like or multipotent characteristics are obtained and wherein the plurality of cells having pluripotent-like or multipotent characteristics are organized within a three-dimensional assembly of cells or a tissue comprising the plurality of cells.

7 . The method of claim 1 , wherein the cell of the first type is selected from the group consisting of an adipose-derived stem cell, a mesenchymal stem cell, a hematopoietic stem cell, a skin derived precursor cell, a hair follicle cell, a fibroblast, a keratinocyte, an epidermal cell, an endothelial cell, an epithelial cell, a granulosa epithelial cell, a melanocyte, an adipocyte, a chondrocyte, a hepatocyte, a B lymphocyte, a T lymphocyte, a granulocyte, a macrophage, a monocyte, a mononuclear cell, a sertoli cell, a neuron, a glial cell, a cardiac muscle cell, and another muscle cell.

8 . The method of claim 7 , wherein the cell of the first cell type is a human fibroblast cell, a human keratinocyte, a human adipose derived stem cell, a human mesenchymal stem cell, or a human hematopoietic stem cell.

9 . The method of claim 1 , further comprising treating the cells of a first cell type with a cytoskeleton disruptor.

10 . The method of claim 9 , wherein the cytoskeleton disruptor is Cytochalasin B or a myosin inhibitor.

11 . The method of claim 1 , wherein the cell having pluripotent-like or multipotent characteristics obtained is capable of differentiating into a unipotent or somatic cell.

Assignments (3)

Continuity (6)

References Cited (139)