Generating induced pluripotent stem cells and progenitor cells from fibroblasts

Patent metadata

Field	Value
Publication number	US-9340772-B2
Application number	US-201013504745-A
Country	US
Kind code	B2
Filing date	Oct 29, 2010
Priority date	Oct 29, 2009
Publication date	May 17, 2016
Grant date	May 17, 2016

Abstract

The present disclosure provides a method of generating progenitor cells, such as hematopoietic or neural progenitor cells, from fibroblasts, such as dermal fibroblasts, comprising providing fibroblasts that express or are treated with a POU domain containing gene or protein and culturing the cells under conditions that allow production of progenitor cells, without traversing the pluripotent state. Also provided is a method of isolating a subpopulation of fibroblasts with reprogramming potential comprising providing fibroblasts that express an Oct-4-reporter and isolating cells that are positive for the reporter. Further provided is a method of generating reprogrammed fibroblast-derived induced pluripotent stem cells. Also provided are uses and assays of the cells produced by the methods of the disclosure.

First claim

The invention claimed is:

1. A method of generating hematopoietic progenitor cells from fibroblasts comprising:
a) providing fibroblasts that express or are treated with a full-length Oct-4 gene or protein; wherein the fibroblasts do not overexpress or ectopically express or are not treated with Sox-2 or Nanog; and
b) culturing the cells of step (a) under conditions that allow production of hematopoietic progenitor cells and isolating CD45+ progenitor cells;
wherein the cells in (b) do not traverse the pluripotent state, and
wherein the fibroblasts of step a) are not treated with any pluripotency factor protein other than said full-length Oct-4 gene or protein.

2. The method of claim 1 , wherein fibroblasts that express the full-length Oct-4 gene or protein in step (a) are produced by lentiviral transduction.

3. The method of claim 1 , wherein fibroblasts treated with the full-length Oct-4 gene or protein comprises providing an exogenous full-length Oct-4 gene or protein.

4. The method of claim 1 , further comprising culturing the cells in step (b) in differentiation medium, wherein the differentiation medium comprises hematopoietic medium comprising at least one hematopoietic cytokine.

5. The method of claim 4 , wherein the at least one hematopoietic cytokine is Flt3 ligand and/or SCF and the differentiated hematopoietic cells are of the myeloblast lineage or wherein the at least one hematopoietic cytokine is EPO and the differentiated hematopoietic cells are of the erythroid or megakaryocytic lineage.

6. The method of claim 1 , wherein the fibroblasts are dermal fibroblasts.

Assignees

• Bhatia Mickie
• Szabo Eva
• Rampalli-Deshpande Shravanti
• Munoz Risueno Ruth
• Univ Mcmaster

Inventors

• Bhatia Mickie
• Szabo Eva
• Rampalli-Deshpande Shravanti
• Munoz Risueno Ruth

Classifications

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N2506/1307

  from adult fibroblasts · CPC title

• C12N2501/602

  Sox-2 · CPC title

• C12N2501/13

  Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins · CPC title