Efficient Generation of Integration-Free Human Induced Pluripotent Stem Cells From Keratinocytes by Simple Transfection of Episomal Vectors

Piao, Yulan; Hung, Stevephen; Lim, Shiang Y.; Wong, Raymond Ching-Bong; Ko, Minoru S.H.

doi:10.5966/sctm.2013-0036

Cited by 52 publications

(48 citation statements)

References 20 publications

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“…We have shown previously that the criteria for using AP expression coupled with hESC morphology represents a reliable method to quantify reprogramming efficiency, with similar accuracy to quantification based on TRA-1-60 expression (Supplementary Fig. 1)20. Figure 5a shows a representative picture of a fully reprogrammed hiPSC colony with hESC morphology and a non-hiPSC colony containing differentiated cells.…”

Section: Resultsmentioning

confidence: 80%

“…This quantification approach has been shown to yield similar numbers of TRA-1-60 positive colonies, demonstrating this as a valid way to identify fully reprogramed hiPSC colonies (Supplementary Fig. 1)20. Subsequently, a clonal hiPSC cell line was isolated from reprogramming from the 2SiPM or 5SiPM substrates.…”

Section: Methodsmentioning

confidence: 82%

“…1 620. Briefly, 50,000 fibroblasts were nucleofected with episomal vectors expressing OCT4, SOX2, KLF4, L-MYC, LIN28 and shRNA against p53 (plasmids were gifts from Prof. Shinya Yamanaka; gene plasmid #27077, #27078, #27080).…”

Section: Methodsmentioning

confidence: 99%

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Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs)

Wang

Hung

Thissen

et al. 2016

Sci Rep

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Human induced pluripotent stem cells (hiPSCs) are capable of differentiating into any cell type and provide significant advances to cell therapy and regenerative medicine. However, the current protocol for hiPSC generation is relatively inefficient and often results in many partially reprogrammed colonies, which increases the cost and reduces the applicability of hiPSCs. Biophysical stimulation, in particular from tuning cell-surface interactions, can trigger specific cellular responses that could in turn promote the reprogramming process. In this study, human fibroblasts were reprogrammed into hiPSCs using a feeder-free system and episomal vectors using novel substrates based on binary colloidal crystals (BCCs). BCCs are made from two different spherical particle materials (Si and PMMA) ranging in size from nanometers to micrometers that self-assemble into hexagonal close-packed arrays. Our results show that the BCCs, particularly those made from a crystal of 2 μm Si and 0.11 μm PMMA particles (2SiPM) facilitate the reprogramming process and increase the proportion of fully reprogrammed hiPSC colonies, even without a vitronectin coating. Subsequent isolation of clonal hiPSC lines demonstrates that they express pluripotent markers (OCT4 and TRA-1-60). This proof-of-concept study demonstrates that cell reprogramming can be improved on substrates where surface properties are tailored to the application.

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Section: Resultsmentioning

confidence: 80%

Section: Methodsmentioning

confidence: 82%

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Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs)

Wang

Hung

Thissen

et al. 2016

Sci Rep

Self Cite

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show abstract

“…Due to the ease of isolation and maintenance for reprogramming, the most popular source is fibroblasts. Other cell types, including keratinocytes (25), mesenchymal stem cells (26), adipose stem cells (27), hair follicular cells (28), neural stem cells (29) and urinary cells (30), have also been used successfully. A more desirable source of starting material is human peripheral blood, which can be readily obtained through non-invasive routine clinical procedures.…”

Section: Induced Pluripotent Stem Cellsmentioning

confidence: 99%

Pluripotent stem cells to hepatocytes, the journey so far

2017

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Abstract. Over the past several years, there has been substantial progress in the field of regenerative medicine, which has enabled new possibilities for research and clinical application. For example, there are ongoing efforts directed at generating functional hepatocytes from adult-derived pluripotent cells for toxicity screening, generating disease models or, in the longer term, for the treatment of liver failure. In the present review, the authors summarise recent developments in regenerative medicine and pluripotent stem cells, the methods and tissues used for reprogramming and the differentiation of induced pluripotent stem cells (iPSCs) into hepatocyte-like cells. In addition, the hepatic disease models developed using iPSC technologies are discussed, as well as the potential for gene editing.

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“…For example, we have used the flowthrough chamber as an effective and comparatively fast model for teratoma formation after implantation of human induced pluripotent stem cells. 41,42 Thus, this approach may be used as a "quality control" tool to achieve tumor-free tissue engineering with pluripotent stem cells. Also, drug toxicology studies could be explored in human tissues grown inside the chamber.…”

mentioning

confidence: 99%

Tissue Engineering by Intrinsic Vascularization in an <em>In Vivo</em> Tissue Engineering Chamber

Zhan

Marré

Mitchell

et al. 2016

JoVE

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In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

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Efficient Generation of Integration-Free Human Induced Pluripotent Stem Cells From Keratinocytes by Simple Transfection of Episomal Vectors

Cited by 52 publications

References 20 publications

Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs)

Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs)

Pluripotent stem cells to hepatocytes, the journey so far

Tissue Engineering by Intrinsic Vascularization in an <em>In Vivo</em> Tissue Engineering Chamber

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