Butyrate Greatly Enhances Derivation of Human Induced Pluripotent Stem Cells by Promoting Epigenetic Remodeling and the Expression of Pluripotency-Associated Genes

Stem Cell Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Stem Cells (Impact Factor: 7.7). 04/2010; 28(4):713-20. DOI: 10.1002/stem.402
Source: PubMed

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.

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Available from: Robert Brodsky, Aug 24, 2015
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    • "iPSCs hold great potential in basic stem cell biology and regenerative medicine (Lowry et al., 2008; Meissner et al., 2007; Park et al., 2008; Takahashi et al., 2007; Yu et al., 2007). Because of the oncogenic features of c-Myc and Klf4 and low efficiency with slow kinetics for iPSC generation, many strategies have been developed to reduce tumorigenesis or enhance reprogramming efficiency (Liao et al., 2011; Mali et al., 2010). microRNAs (miRNAs) are 18-to 24-nucleotide singlestranded RNAs that can degrade target mRNAs or repress the translation of encoded proteins by binding to partially complementary target sites (Bartel, 2004). "
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    ABSTRACT: Abstract Small-molecule inhibitors and microRNAs (miRNAs) are two newly emerging classes of tools for optimizing induced pluripotent stem cell (iPSC) generation. We report here that sodium butyrate (NaB), a small-molecule inhibitor of histone deacetylases (HDACs), upregulates transcriptional levels of the miR-302/367 cluster by enhancing Oct4 transcriptional activity at the miR-302/367 cluster promoter. NaB does not affect the OCT4 DNA-binding domain; instead it enhances transactivity of the OCT4 transactivation domains. We elucidate that OCT4 transcriptional activity is usually dampened by its associated HDACs in cells and can be derepressed by NaB by impairing the interaction between Oct4 and HDACs, which leads to an elevated expression of the miR-302/367 cluster. Our new findings suggest a novel molecular mechanism for NaB in promoting somatic cell reprogramming via the miR-302/367 cluster.
    02/2014; 16. DOI:10.1089/cell.2013.0070
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    • "Among them, sodium butyrate, a histone deacetylase inhibitor, is the most important one for reprogramming of blood cells. Butyrate substantially enhances iPSC derivation from both mouse and human cells by increasing the expression of endogenous pluripotency-associated genes and upregulating the expression of miR-302/367 cluster [49] [50] [51]. Other factors such as NANOG [52], LIN28 [53], and miR- 302 family [54] also play important roles in boosting reprogramming efficiency. "
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    ABSTRACT: Breakthroughs in cell fate conversion have made it possible to generate large quantities of patient-specific cells for regenerative medicine. Due to multiple advantages of peripheral blood cells over fibroblasts from skin biopsy, the use of blood mononuclear cells (MNCs) instead of skin fibroblasts will expedite reprogramming research and broaden the application of reprogramming technology. This review discusses current progress and challenges of generating induced pluripotent stem cells (iPSCs) from peripheral blood MNCs and of in vitro and in vivo conversion of blood cells into cells of therapeutic value, such as mesenchymal stem cells, neural cells and hepatocytes. An optimized design of lentiviral vectors is necessary to achieve high reprogramming efficiency of peripheral blood cells. More recently, non-integrating vectors such as Sendai virus and episomal vectors have been successfully employed in generating integration-free iPSCs and somatic stem cells.
    09/2013; 11(5). DOI:10.1016/j.gpb.2013.09.001
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    • "Research article variety of different reprogramming factors, reprogramming enhancers, and cell types have been evaluated (Liao et al., 2008; Judson et al., 2009; Mali et al., 2010; Cheng et al., 2011; Niibe et al., 2011; Szablowska-Gadomska et al., 2011; Zhang et al., 2011; Li and Rana, 2012; Lin et al., 2012; Liu et al., 2012; Mahmoudi and Brunet, 2012; Okita et al., 2013; Zhang and Wu, 2013). Of the accessible cell types used for iPSC generation, the reprogramming of keratinocytes has been shown to be as much as 100-fold more efficient and at least twofold faster than the reprogramming of dermal fibroblasts (Aasen et al., 2008). "
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    ABSTRACT: Next-generation and Sanger sequencing were combined to identify disease-causing USH2A mutations in an adult patient with autosomal recessive RP. Induced pluripotent stem cells (iPSCs), generated from the patient’s keratinocytes, were differentiated into multi-layer eyecup-like structures with features of human retinal precursor cells. The inner layer of the eyecups contained photoreceptor precursor cells that expressed photoreceptor markers and exhibited axonemes and basal bodies characteristic of outer segments. Analysis of the USH2A transcripts of these cells revealed that one of the patient’s mutations causes exonification of intron 40, a translation frameshift and a premature stop codon. Western blotting revealed upregulation of GRP78 and GRP94, suggesting that the patient’s other USH2A variant (Arg4192His) causes disease through protein misfolding and ER stress. Transplantation into 4-day-old immunodeficient Crb1−/− mice resulted in the formation of morphologically and immunohistochemically recognizable photoreceptor cells, suggesting that the mutations in this patient act via post-developmental photoreceptor degeneration. DOI:
    eLife Sciences 08/2013; 2:e00824. DOI:10.7554/eLife.00824 · 8.52 Impact Factor
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