Feeder‐Free Reprogramming of Human Fibroblasts with Messenger RNA. (13th November 2013)

• Record Type: Journal Article
• Authors: Warren, Luigi; Wang, Jiwu
• Editors: Bhatia, Mick; Elefanty, Andrew G.; Fisher, Susan J.; Patient, Roger; Schlaeger, Thorsten; Snyder, Evan Y.
• Is Part Of: Current protocols in stem cell biology. Volume 27(2013)
• Page Start: 4A.6.1
• Page End: 4A.6.27
• Publication Date: 2013-11-13
• Subjects: induced pluripotent stem cells -- iPSC -- mRNA reprogramming -- feeder‐free -- xeno‐free; Stem cells -- Laboratory manuals; Stem cells -- Physiology; Stem Cells; Stem cells; Laboratory Manual; Laboratory manuals; 571.5
• DOI: 10.1002/9780470151808.sc04a06s27
• Languages: English
• ISSNs: 1941-7322

Abstract

Abstract: This unit describes a feeder‐free protocol for deriving induced pluripotent stem cells (iPSCs) from human fibroblasts by transfection of synthetic mRNA. The reprogramming of somatic cells requires transient expression of a set of transcription factors that collectively activate an endogenous gene regulatory network specifying the pluripotent phenotype. The necessary ectopic factor expression was first effected using retroviruses; however, as viral integration into the genome is problematic for cell therapy applications, the use of footprint‐free vectors such as mRNA is increasingly preferred. Strong points of the mRNA approach include high efficiency, rapid kinetics, and obviation of a clean‐up phase to purge the vector. Still, the method is relatively laborious and has, up to now, involved the use of feeder cells, which brings drawbacks including poor applicability to clinically oriented iPSC derivation. Using the methods described here, mRNA reprogramming can be performed without feeders at much‐reduced labor and material costs relative to established protocols. Curr. Protoc. Stem Cell Biol . 27:4A.6.1‐4A.6.27. © 2013 by John Wiley & Sons, Inc.