Generation of patient-specific iPSC
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 in definitely 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 three reprogramming methods based on the delivery of the reprogramming factors. These include non-integrating methods such as Sendai-based and Episomal-based delivery systems. The table below summarizes all the relevant information regarding the three methodologies.
|Reprogramming Method||Efficiency||Cost for NU Researchers||Time||Integration free||Cell Type|
|CytoTune®-iPS 2.0 Sendai Reprogramming Kit||0.02 – 1.2%||$2,000||~3 months||Yes||Skin fibroblast and PBMC|
|Epi5™ Episomal iPSC Reprogramming Kit||0.04–0.3%||$2,000||~3 months||Yes||Skin fibroblast|
|Retrovirus Reprogramming||<1%||$2,000||~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 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.