title: Feeder-free derivation of human induced pluripotent stem cells with messenger RNA.
abstract: The therapeutic promise of induced pluripotent stem cells (iPSCs) has spurred efforts to circumvent genome alteration when reprogramming somatic cells to pluripotency. Approaches based on episomal DNA, Sendai virus, and messenger RNA (mRNA) can generate "footprint-free" iPSCs with efficiencies equaling or surpassing those attained with integrating viral vectors. The mRNA method uniquely affords unprecedented control over reprogramming factor (RF) expression while obviating a cleanup phase to purge residual traces of vector. Currently, mRNA-based reprogramming is relatively laborious due to the need to transfect daily for ~2 weeks to induce pluripotency, and requires the use of feeder cells that add complexity and variability to the procedure while introducing a route for contamination with non-human-derived biological material. We accelerated the mRNA reprogramming process through stepwise optimization of the RF cocktail and leveraged these kinetic gains to establish a feeder-free, xeno-free protocol which slashes the time, cost and effort involved in iPSC derivation.
authors.affiliation: Allele Biotechnology & Pharmaceuticals, Inc, San Diego, CA, USA.
authors.forename: Luigi
authors.initials: L
authors.lastname: Warren
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Ni
authors.forename: Jiwu
authors.initials: J
authors.lastname: Wang
authors.forename: Xirong
authors.initials: X
authors.lastname: Guo
mesh_terms: D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D017570:MyoD Protein / Q000235:genetics; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D012333:RNA, Messenger / Q000502:physiology*
chemical_list: D017405:Herpes Simplex Virus Protein Vmw65; D018398:Homeodomain Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012333:RNA, Messenger