Generation of mouse-induced pluripotent stem cells by transient expression of a single nonviral polycistronic vector.
Induced pluripotent stem (iPS) cells have generated keen interest due to their potential use in regenerative medicine. They have been obtained from various cell types of both mice and humans by exogenous delivery of different combinations of Oct4, Sox2, Klf4, c-Myc, Nanog, and Lin28. The delivery of these transcription factors has mostly entailed the use of integrating viral vectors (retroviruses or lentiviruses), carrying the risk of both insertional mutagenesis and oncogenesis due to misexpression of these exogenous factors. Therefore, obtaining iPS cells that do not carry integrated transgene sequences is an important prerequisite for their eventual therapeutic use. Here we report the generation of iPS cell lines from mouse embryonic fibroblasts with no evidence of integration of the reprogramming vector in their genome, achieved by nucleofection of a polycistronic construct coexpressing Oct4, Sox2, Klf4, and c-Myc.
Gonzalez
Federico
F
Center of Regenerative Medicine of Barcelona, Dr. Aiguader 88, p7, Barcelona 08003, Spain.
Barragan Monasterio
Montserrat
M
Tiscornia
Gustavo
G
Montserrat Pulido
Nuria
N
Vassena
Rita
R
Batlle Morera
Laura
L
Rodriguez Piza
Ignasi
I
Izpisua Belmonte
Juan Carlos
JC
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012038:Regeneration; D014157:Transcription Factors / Q000235:genetics*
C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors