title: Genetic manipulation of human induced pluripotent stem cells.
abstract: Human induced pluripotent stem cells (HIPSC) have tremendous value as a source of autologous cells for cellular transplantation in the treatment of degenerative diseases. The protocols described here address methods for large-scale genetic modification of HIPSCs. The first is an optimized method for transfecting HIPSCs cultured in feeder-free conditions. The second method allows nucleofection of trypsinized HIPSCs at an optimal cell density. Both methods enable robust generation of stable HIPSC transfectants within two weeks. Our protocols are highly reproducible and do not require optimization for individual HIPSC and human embryonic stem cell (HESC) lines.
authors.affiliation: Stem Cell Core, Department of Cell Biology and Neuroscience, University of California Riverside, Riverside, California, USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Wang
authors.forename: Chee Gee
authors.initials: CG
authors.lastname: Liew
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism; D061252:Feeder Cells / Q000166:cytology / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014162:Transfection