A protocol describing the genetic correction of somatic human cells and subsequent generation of IPS cell

Rodríguez Pizà, Ignasi; Raya Chamorro, Ángel; Navarro, Susanna; Izpisúa Belmonte, J. C.; Bueren, Juan; Consiglio, Antonella; Sánchez Danés, Adriana, 1984-; Guenechea, Guillermo; Richaud Patin, Yvonne

doi:http://dx.doi.org/10.1038/nprot.2010.9

A protocol describing the genetic correction of somatic human cells and subsequent generation of IPS cell

Citation

Raya Á, Rodríguez I, Navarro S, Richaud-Patin Y, Guenechea G, Sánchez-Danés A et al. A protocol describing the genetic correction of somatic human cells and subsequent generation of iPS cells. Nat Protoc. 2010;5:647-60. DOI 10.1038/nprot.2010.9

Abstract

The generation of patient-specific induced pluripotent stem cells (iPSCPSCPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSCPSCPSC technology can be used to generate disease-free progenitor cells of potential interest for autologous cell therapy. We explain a protocol for the reproducible generation of genetically corrected iPSCPSCPSCs starting from the skin biopsies of Fanconi anemia patients using retroviral transduction with OCT4, SOX2 and KLF4. Before reprogramming, the fibroblasts and/or keratinocytes of the patients are genetically corrected with lentiviruses expressing FANCA. The same approach may be used for other diseases susceptible to gene therapy correction. Genetically corrected, characterized lines of patient-specific iPSCPSCPSCs can be obtained in 4–5 months.