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Faster and Safer Method for Differentiation of Human Stem Cells

Method also allows for unlimited expansion potential of stem cells without the use of feeder cells.  

INVENTION: Investigators at the Salk Institute have developed an efficient method for the directed differentiation of human embryonic stem cells (hES) and human induced pluripotent stem cells (iPSC) into vascular progenitor cells. By analyzing well known mediators of mesodermal development in different human pluripotent stem cell (hPSC) lines, researchers were able to develop a mesodermal induction medium that can efficiently differentiate hPSCs into vascular progenitor cells. Multipotent progenitor cells can be generated after 8 days using the mesodermal induction medium without the use of feeder cells, and are able to generate functional, differentiated cells of the endothelial, smooth muscle and hematopoietic lineages. Because of the short differentiation time and potential for unlimited expansion, this new method has enormous potential for advancing the development of new drug and cell regeneration therapies. 

Figure: a) Schematic representation of the conversion process towards CD34+ progenitor cells and further differentiation into terminally differentiated endothelial cells. b) Representative flow cytometry plots demonstrating absent expression of pluripotency-associated markers upon plastic induction with non-integrative plasmids followed by Mesodermal Induction Media (MIM) differentiation. c) Representative flow cytometry analysis of CD34 expression before and after MIM differentiation in human fibroblasts (BJ) induced to a plastic state by the use of non-integrative approaches in the presence of mi302-367. d) Representative flow cytometry quantification of BJ-converted VE-cadherin+ and endoglin+ endothelial cells derived in the presence of mi302/367. e) mRNA expression profile showing specific upregulation of endothelial markers. f) Fluorescence microscopy analysis showing the expression of the indicated endothelial markers in converted cells. g) Characterization of endothelial subtypes in BJ converted endothelial cells. h) Representative pictures of endothelial cells derived by non-integrative approaches upon LDL uptake. i) BJ-derived endothelial cells converted by non-integrative approaches spontaneously formed capillary-like structures.

APPLICATIONS:

  • Generation of endothelial cells, smooth muscle cells, and monocytic and erythroid lineages of hematopoietic cells 
  • Study and treatment of ischemic pathologies
  • Identification of novel drug targets and biomarkers
  • Generation of relevant cell types for mechanism of action studies
  • Cell regeneration therapies

ADVANTAGES

  • Highly efficient method
  • Unlimited expansion potential of the stem cells
  • Short differentiation time 
  • Reduced risk of genetic alterations due to short differentiation time
  • Vascular progenitor cells can be efficiently differentiated into functional cells

STAGE OF DEVELOPMENT:   In vitro and  in vivo data available; animal studies are underway.

BACKGROUND: Multipotent vascular progenitor cells hold great potential for clinical applications. Current methods used to generate multipotent progenitor cells have many limitations including inefficient differentiation, limited expansion potential, purity issues, long differentiation times and the use of feeder cells. This new method overcomes many of the barriers that have limited the progression of stem cell therapies for cell regeneration.

INVENTORS: Juan Carlos Izpisua Belmonte, Leo Kurian, Emmanuel Nivet, and Ignacio Sancho-Martnez
PATENT STATUS: PCT Patent Application No. PCT/US2012/025449 
PUBLICATION: Nature Methods 2013 Jan:10(1):77-83.
CONTACT: Melissa J. Rodgers, Ph.D., 858.453.4100 x1481, mrodgers@salk.edu
REFERENCE #: S11001
Patent Information:
For Information, Contact:
Michelle Booden
Director, Licensing and Intellectual Property
Salk Institute
mbooden@salk.edu
Inventors:
Keywords:
Belmonte
Canine Stem Cells
Cardiovascular Disease
Cell Culture Media
Cell Lines
Equine Stem Cells
Ischemia
Regenerative Medicine
Reprogramming
Stem Cells
Vascular disease
Vascular progenitor cells
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