iPSC

Induced Pluripotent Stem Cells (iPSCs)

Initially described in the pioneering work of Yamanaka and colleagues, the ability to "reprogram" differentiated somatic cells into a pluripotent embryonic stem cell-like state by retroviral mediated expression of four specific transcription factors has revolutionized our ability to develop new models to study human disease and represents a significant step towards patient-specific cell replacement therapies.

What can I do with iPSCs?

In addition to solving ethical concerns related to the use of blastocyst-derived embryonic stem cells, the use of iPSCs for the generation of therapeutic cells for cell replacement therapy may avoid the requirement for post-transplant immune suppression because iPSCs can be generated directly from the transplant recipient and will therefore be genetically identical to the patient. Additionally, because it is possible to reprogram somatic cells derived from diseased individuals iPSC technology provides an important new platform for the development of new models of human disease. Thus, upon appropriate differentiation these cells can then be used to study normal and pathologic human tissue development in vitro, enabling new insights into disease pathology as well as the development of novel therapeutic agents and patient-specific cell replacement therapies.
 

 
 

What has the PSCF done with iPSCs?

Using the original "Yamanaka cocktail" of transcription factors the PSCF has successfully generated iPSCs from human keratinocytes, fibroblasts, bone marrow and cord blood cells. These iPSCs have typical hESC-like colony morphology and express pluripotency markers (e.g. Tra-1-81, Dnmt3b).
 
 
Functional pluripotency of our human iPSC cells is determined using teratoma assay. When injected sub-cutaneously in immune-compromised mice, pluripotent cells differentiate into benign tumors containing histologically-recognisable tissues derived from all three primary germ layers (endoderm, ectoderm and mesoderm). Hematoxlyin and eosin stained sections derived from a keratinocyte-derived iPSC line are shown. Specific endoderm, ectoderm and mesoderm-derived tissues are indicated by arrows.
 
 
Additionally, we have efficiently differentiated these iPSC lines into FoxA2/Sox17 positive definitive endoderm using a published directed differentiation procedure (D'Amour et al., Nat Biotech 2005). Analyses of additional directed differentiation protocols is underway.
 

 

iPSC Services

The PSCF is committed to facilitating access to this exciting technology for CCHMC/UC researchers. We can provide the following iPSC services:

  • Training in iPSC generation
  • Reprogramming factor expression and generation of reprogrammed colonies
  • Expansion of reprogrammed colonies
  • Pluripotency characterization
  • Banking of iPSCs

Our iPSC services range from training in iPSC generation to complete iPSC generation service
Please contact us to discuss the details and costs associated with these services

Notes

  • Currently we are using the "Yamanaka" reprogramming factor cocktail: Oct4, Sox2, Klf4 and cMyc expressed from pMXs retroviral vectors
  • However, we are testing new reprogramming approaches as well as working with CCHMC investigators to develop and characterize improved vectors for iPSC generation 
  • The PSCF utilizes the CCHMC viral vector core for generation of recombinant viruses for iPSC generation