Format

Send to

Choose Destination
Nat Protoc. 2010 Apr;5(4):647-60. doi: 10.1038/nprot.2010.9. Epub 2010 Mar 11.

A protocol describing the genetic correction of somatic human cells and subsequent generation of iPS cells.

Author information

1
Center for Regenerative Medicine in Barcelona, Barcelona, Spain.

Abstract

The generation of patient-specific induced pluripotent stem cells (iPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSC 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 iPSCs 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 iPSCs can be obtained in 4-5 months.

PMID:
20224565
DOI:
10.1038/nprot.2010.9
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Nature Publishing Group
Loading ...
Support Center