Derivation of hiPS cells
The simultaneous development of methods for reprogramming adult cells into induced pluripotent stem cells (iPSCs) (Park et al, 2008; Takahashi et al, 2007; Yu et al, 2007) and the directed differentiation of pluripotent stem cells into distinct somatic cell types, suggested an attractive route to a novel approach for developing disease model systems. Patient-specific iPSCs can be generated by epigenetic reprogramming of various adult cell types such as skin fibroblasts and blood mononuclear cells and just like embryonic stem cells (ESCs), self-renew indefinitely and retain the potential to give rise to all cell types in the human body (Takahashi et al, 2007).
At the Stem Cell Core Facility we provide investigators with custom iPSC lines and offer Sendai-based non-integrating method of reprogramming of adult dermal skin fibroblasts or Peripheral Blood Mononuclear cells. If the nature of the research project requires a different method of iPSC reprogramming please discuss this with us and we will review other options.
|Reprogramming Method||Efficiency %||Cost for NU Researchers||Cost for NINDS-NU Researchers||Time||Integration Fee||Cell Type
|CytoTune® iPS 2.0 Sendai Reprogramming Kit||0.02 - 1/2||$1,000||$600||2-3 months||Yes||Skin Fibroblast and PBMC (Peripheral Blood Mononuclear Cells)|
|Epi5™ Episomal iPSC Repgrogramming Kit||0.04 - 0.3||$1,000||$600||2-3 months||Yes||Skin Fibroblast|
|Retrovirus Reprogramming||<1||$950||$475||2-3 months||No||Skin Fibroblast|
Once an iPSC line is generated we test it for mycoplasma and provide investigators with two frozen vials. We highly recommend appropriate quality control (QC) of all newly derived iPSC lines. This can be done at the core facility (for an additional charge) or by the investigators themselves. The QC that we recommend consists of (a) immunostaining for the pluripotency markers, e.g., NANOG/SSEA4, (b) karyotype analysis and (c) the Scorecard Assay.* We would be happy to discuss these options further and help you make the right choice for your specific research project.
*Dr. Kiskinis is one of the inventors of the Scorecard Assay and holds a US Patent on it