Reprogramming of cells to a new fate

Patent metadata

Field	Value
Publication number	US-9556417-B2
Application number	US-201615072769-A
Country	US
Kind code	B2
Filing date	Mar 17, 2016
Priority date	Jun 14, 2010
Publication date	Jan 31, 2017
Grant date	Jan 31, 2017

Abstract

The present invention generally provides methods and compositions for transdifferentiation of an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate. Also provided are methods and compositions for the transdifferentiation of an animal cell from a non-pluripotent mesodermal, endodermal, or ectodermal cell fate to a different non-pluripotent mesodermal, endodermal, or ectodermal cell fate.

First claim

What is claimed is:

1. A method of converting an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate, the method comprising:
(a) introducing a polynucleotide encoding an Oct4 polypeptide, and optionally one or more reprogramming factors comprising polynucleotides encoding a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide, into a first non-pluripotent cell, or contacting a first non-pluripotent cell with an Oct4 polypeptide, and optionally one or more reprogramming factors comprising a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide,
(b) limiting the expression of endogenous Nanog in the cells from step (a) to a level substantially lower than the level of expression of endogenous Nanog in an induced pluripotent cell (iPSC), thereby generating a non-pluripotent intermediate cell and
(c) inducing differentiation of the cell under conditions to generate a cell having a second non-pluripotent cell fate,
wherein the first non-pluripotent cell is a mesodermal, ectodermal or endodermal cell, and wherein the cell having the second non-pluripotent cell fate from step (c) is in the same or different cell lineage as the first non-pluripotent cell.

2. The method of claim 1 , wherein step (b) comprises eliminating exogenous LIF.

3. The method of claim 1 , wherein the first non-pluripotent cell is a fibroblast or a neural precursor cell.

4. The method of claim 1 , wherein inducing differentiation of the non-pluripotent intermediate cell to the cell having the second non-pluripotent cell fate comprises contacting the non-pluripotent intermediate cell with BMP4, a calcium channel agonist, a Gas activating agent, a cAMP analog, and/or a GSK-3 inhibitor.

5. The method of claim 1 , wherein the method is conducted in vitro.

Assignees

• Scripps Research Inst

Inventors

• Efe Jem A
• Kim Janghwan
• Zhu Saiyong
• Hilcove Simon
• Ding Sheng

Classifications

• C12N5/0623 Primary

  Stem cells · CPC title

• C12N2501/602

  Sox-2 · CPC title

• A61K35/30

  Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue · CPC title

• C12N5/0678

  Stem cells; Progenitor cells; Precursor cells · CPC title

• C12N2501/727

  Kinases (EC 2.7.) · CPC title