title: Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes.
abstract: We report here that butyrate, a naturally occurring fatty acid commonly used as a nutritional supplement and differentiation agent, greatly enhances the efficiency of induced pluripotent stem (iPS) cell derivation from human adult or fetal fibroblasts. After transient butyrate treatment, the iPS cell derivation efficiency is enhanced by 15- to 51-fold using either retroviral or piggyBac transposon vectors expressing 4 to 5 reprogramming genes. Butyrate stimulation is more remarkable (>100- to 200-fold) on reprogramming in the absence of either KLF4 or MYC transgene. Butyrate treatment did not negatively affect properties of iPS cell lines established by either 3 or 4 retroviral vectors or a single piggyBac DNA transposon vector. These characterized iPS cell lines, including those derived from an adult patient with sickle cell disease by either the piggyBac or retroviral vectors, show normal karyotypes and pluripotency. To gain insights into the underlying mechanisms of butyrate stimulation, we conducted genome-wide gene expression and promoter DNA methylation microarrays and other epigenetic analyses on established iPS cells and cells from intermediate stages of the reprogramming process. By days 6 to 12 during reprogramming, butyrate treatment enhanced histone H3 acetylation, promoter DNA demethylation, and the expression of endogenous pluripotency-associated genes, including DPPA2, whose overexpression partially substitutes for butyrate stimulation. Thus, butyrate as a cell permeable small molecule provides a simple tool to further investigate molecular mechanisms of cellular reprogramming. Moreover, butyrate stimulation provides an efficient method for reprogramming various human adult somatic cells, including cells from patients that are more refractory to reprogramming.
authors.affiliation: Stem Cell Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Prashant
authors.initials: P
authors.lastname: Mali
authors.forename: Bin-Kuan
authors.initials: BK
authors.lastname: Chou
authors.forename: Jonathan
authors.initials: J
authors.lastname: Yen
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Ye
authors.forename: Jizhong
authors.initials: J
authors.lastname: Zou
authors.forename: Sarah
authors.initials: S
authors.lastname: Dowey
authors.forename: Robert A
authors.initials: RA
authors.lastname: Brodsky
authors.forename: Joyce E
authors.initials: JE
authors.lastname: Ohm
authors.forename: Wayne
authors.initials: W
authors.lastname: Yu
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Baylin
authors.forename: Kosuke
authors.initials: K
authors.lastname: Yusa
authors.forename: Allan
authors.initials: A
authors.lastname: Bradley
authors.forename: David J
authors.initials: DJ
authors.lastname: Meyers
authors.forename: Chandrani
authors.initials: C
authors.lastname: Mukherjee
authors.forename: Philip A
authors.initials: PA
authors.lastname: Cole
authors.forename: Linzhao
authors.initials: L
authors.lastname: Cheng
mesh_terms: D001483:Base Sequence; D002087:Butyrates / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D016922:Cellular Senescence; D004251:DNA Transposable Elements; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D016415:Sequence Alignment
chemical_list: D002087:Butyrates; D004251:DNA Transposable Elements; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4