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    Establishment of SIAISi018-A, an induced pluripotent stem cell (iPSC) line from a healthy 45-year-old Chinese Han
    Stem Cell Research (2022)
    Yi ZhangJiajun ShouWentao GuWeihao DiJian Zhao
    A 45-year-old Han Male from China contributed peripheral blood mononuclear cells induced by reprogramming human OKSM transcription factors (OCT3/4, KLF4 SOX2 and C-MYC) with a non-integrated additional vector system. Immunological markers confirmed the pluripotent nature of IPSC. Spontaneous tridermal differentiation confirmed the differentiation ability of IPSC with normal karyotype.
    Reprogramming
    KLF4
    Citations (0)
    Embryonic stem cell markers expression in cancers
    Biochemical and Biophysical Research Communications (2009)
    Matthieu SchoenhalsAlboukadel KassambaraJohn De VosDirk HoseJérôme Moreaux
    KLF4
    Homeobox protein NANOG
    Citations (241)
    Metadichol, A natural ligand for expression of Yamanaka reprogramming factors in somatic and primary cancer cell lines
    Research Square (Research Square) (2022)
    P.R. Raghavan
    Abstract The conversion of somatic cells back into induced pluripotent stem cells (iPSCs) or embryonic-like stem cells involves the introduction of four genes commonly called Yamanaka factors, i.e., Sox2, Oct4, Klf4, and c-Myc, into the cells. Because these genes and the viral vectors used to introduce them into cells have the potential to cause cancer, iPSC lines are not clinically useful. Most instances of direct reprogramming have achieved have been by the forced expression of defined factors using viral vectors. Here, we show that Metadichol® has the potential to generate iPSCs non-virally and may be helpful in clinical applications. Metadichol is a nanoformulation of long-chain alcohols derived from food. Quantitative real-time PCR (qRT-PCR) and western blotting showed that OCT4, SOX2, and Nanog are expressed when fibroblasts were treated with Metadichol at one picogram to 100 nanograms. Reverse-transcription PCR (RT-PCR) also revealed that OCT4, KLF4, Nanog, and Sox2 levels increased compared to controls by 4.01-, 3.51- and 1.26-, and 2.5-fold, respectively, in A549 cancer cells. In Colo-205 cells, OCT4, KLF4, and Sox2 were increased by 1.79-, 13.17-, and 2.25-folds, respectively. Metadichol treatment with triple-negative primary breast cancer (HCAF-TNPBC) primary cancer cells led to multifold increases of OKSM factors by 19-, 6-, 8.07-, 2.45-, and 6.91-folds in concentration ranges 1picogram to 100 nanograms. Metadichol is a natural product that induces the expression of Yamanaka factors needed for reprogramming and Klotho, an anti-aging gene, and curbs the expression of the TP53 gene, which is critical for reprogramming somatic cells into IPSCs.Metadichol increases endogenous vitamin C levels, leading to the efficient reprogramming of somatic cells into iPSCs. Metadichol is nontoxic and commercially available as a nutritional supplement. Thus, it can be directly tested in vivo in human subjects to confirm that cells can indeed be programmed into a state of induced pluripotency and cause the mitigation of disease conditions.
    KLF4
    Homeobox protein NANOG
    Reprogramming
    Citations (0)
    Generation and characterization of three induced pluripotent stem cell lines (NUIGi046-A, NUIGi046-B, NUIGi046-C) from a 51-year-old healthy individual
    Stem Cell Research (2021)
    Min LiuNing GeYong-Feng HanJamie ReillyMeimei Yang
    Skin punch biopsy was donated by a healthy 51-year-old Caucasian male and the dermal fibroblasts were reprogrammed into human induced pluripotent stem cell (hiPSC) lines by using non-integrative Sendai viruses expressing OCT4, SOX2, KLF4 and c-MYC. Three iPSC lines (NUIGi046-A, NUIGi046-B, NUIGi046-C) highly expressed the pluripotent markers and were capable of differentiating into cells of endodermal, mesodermal, and ectodermal origin. These iPSCs can be offered as controls and in combination with genome-editing and three-dimensional (3D) system. They may be used for human disease modelling and drug screening.
    KLF4
    Sendai virus
    Citations (0)
    Two-Phase Analysis of Molecular Pathways Underlying Induced Pluripotent Stem Cell Induction
    Stem Cells (2011)
    Zhaoyu LinPhilip L. PérezDebin LeiJingyue XuXiang Gao
    Induced pluripotent stem cells (iPSCs) can be reprogrammed from adult somatic cells by transduction with Oct4, Sox2, Klf4, and c-Myc, but the molecular cascades initiated by these factors remain poorly understood. Impeding their elucidation is the stochastic nature of the iPS induction process, which results in heterogeneous cell populations. Here we have synchronized the reprogramming process by a two-phase induction: an initial stable intermediate phase following transduction with Oct4, Klf4, and c-Myc, and a final iPS phase following overexpression of Sox2. This approach has enabled us to examine temporal gene expression profiles, permitting the identification of Sox2 downstream genes critical for induction. Furthermore, we have validated the feasibility of our new approach by using it to confirm that downregulation of transforming growth factor β signaling by Sox2 proves essential to the reprogramming process. Thus, we present a novel means for dissecting the details underlying the induction of iPSCs, an approach with significant utility in this arena and the potential for wide-ranging implications in the study of other reprogramming mechanisms.
    KLF4
    Reprogramming
    Citations (15)
    Metadichol, A natural ligand for expression of Yamanaka reprogramming factors in somatic and primary cancer cell lines
    Research Square (Research Square) (2022)
    P.R. Raghavan
    Abstract The conversion of somatic cells back into induced pluripotent stem cells (iPSCs) or embryonic-like stem cells involves the introduction of four genes commonly called Yamanaka factors, i.e., Sox2, Oct4, Klf4, and c-Myc, into the cells. Because these genes and the viral vectors used to introduce them into cells have the potential to cause cancer, iPSC lines are not clinically useful. Most instances of direct reprogramming have achieved have been by the forced expression of defined factors using viral vectors. Here, we show that Metadichol® has the potential to generate iPSCs non-virally and may be helpful in clinical applications. Metadichol is a nanoformulation of long-chain alcohols derived from food. Quantitative real-time PCR (qRT-PCR) and western blotting showed that OCT4, SOX2, and Nanog are expressed when fibroblasts were treated with Metadichol at one picogram to 100 nanograms. Reverse-transcription PCR (RT-PCR) also revealed that OCT4, KLF4, Nanog, and Sox2 levels increased compared to controls by 4.01, 3.51 and 1.26, and 2.5-fold, respectively, in A549 cancer cells. In Colo-205 cells, OCT4, KLF4, and Sox2 were increased by 1.79, 13.17, and 2.25folds, respectively. Metadichol treatment with triple-negative primary breast cancer (HCAF-TNPBC) primary cancer cells led to multifold increases of OKSM factors by 19.6, 8.07, 2.45, and 6.91, folds increase in Metadichol concentration ranges one picogram to 100 nano grams. Metadichol is a natural product that induces the expression of Yamanaka factors needed for reprogramming and Klotho, an anti-aging gene, and curbs the expression of the TP53 gene, which is critical for reprogramming somatic cells into IPSCs.Metadichol increases endogenous vitamin C levels, leading to the efficient reprogramming of somatic cells into iPSCs. Metadichol is nontoxic and commercially available as a nutritional supplement. Thus, it can be directly tested in vivo in human subjects to confirm that cells can indeed be programmed into a state of induced pluripotency and cause the mitigation of disease conditions.
    KLF4
    Homeobox protein NANOG
    Reprogramming
    Citations (4)
    Metadichol, A natural ligand for expression of Yamanaka reprogramming factors in somatic and primary cancer cell lines
    Research Square (Research Square) (2022)
    P.R. Raghavan
    Abstract The conversion of somatic cells back into induced pluripotent stem cells (iPSCs) or embryonic-like stem cells involves the introduction of four genes commonly called Yamanaka factors, i.e., Sox2, Oct4, Klf4, and c-Myc, into the cells. Because these genes and the viral vectors used to introduce them into cells have the potential to cause cancer, iPSC lines are not clinically useful. Most instances of direct reprogramming have achieved have been by the forced expression of defined factors using viral vectors. Here, we show that Metadichol® has the potential to generate iPSCs non-virally and may be helpful in clinical applications. Metadichol is a nanoformulation of long-chain alcohols derived from food. Quantitative real-time PCR (qRT-PCR) and western blotting showed that OCT4, SOX2, and Nanog are expressed when fibroblasts were treated with Metadichol at one picogram to 100 nanograms. Reverse-transcription PCR (RT-PCR) also revealed that OCT4, KLF4, Nanog, and Sox2 levels increased compared to controls by 4.01, 3.51 and 1.26, and 2.5-fold, respectively, in A549 cancer cells. In Colo-205 cells, OCT4, KLF4, and Sox2 were increased by 1.79, 13.17, and 2.25folds, respectively. Metadichol treatment with triple-negative primary breast cancer (HCAF-TNPBC) primary cancer cells led to multifold increases of OKSM factors by 19, 6, 8.07, 2.45, and 6.91folds in concentration ranges one picogram to 100 nano grams. Metadichol is a natural product that induces the expression of Yamanaka factors needed for reprogramming and Klotho, an anti-aging gene, and curbs the expression of the TP53 gene, which is critical for reprogramming somatic cells into IPSCs.Metadichol increases endogenous vitamin C levels, leading to the efficient reprogramming of somatic cells into iPSCs. Metadichol is nontoxic and commercially available as a nutritional supplement. Thus, it can be directly tested in vivo in human subjects to confirm that cells can indeed be programmed into a state of induced pluripotency and cause the mitigation of disease conditions.
    KLF4
    Homeobox protein NANOG
    Reprogramming
    Citations (0)
    Optimal Ratio of Transcription Factors for Somatic Cell Reprogramming
    Journal of Biological Chemistry (2012)
    Go NagamatsuShigeru SaitoTakeo KosakaKeiyo TakuboTaisuke Kinoshita
    Somatic cell reprogramming is achieved by four reprogramming transcription factors (RTFs), Oct3/4, Sox2, Klf4, and c-Myc. However, in addition to the induction of pluripotent cells, these RTFs also generate pseudo-pluripotent cells, which do not show Nanog promoter activity. Therefore, it should be possible to fine-tune the RTFs to produce only fully pluripotent cells. For this study, a tagging system was developed to sort induced pluripotent stem (iPS) cells according to the expression levels of each of the four RTFs. Using this system, the most effective ratio (Oct3/4-high, Sox2-low, Klf4-high, c-Myc-high) of the RTFs was 88 times more efficient at producing iPS cells than the worst effective ratio (Oct3/4-low, Sox2-high, Klf4-low, c-Myc-low). Among the various RTF combinations, Oct3/4-high and Sox2-low produced the most efficient results. To investigate the molecular basis, microarray analysis was performed on iPS cells generated under high (Oct3/4-high and Sox2-low) and low (Oct3/4-low and Sox2-high) efficiency reprogramming conditions. Pathway analysis revealed that the G protein-coupled receptor (GPCR) pathway was up-regulated significantly under the high efficiency condition and treatment with the chemokine, C-C motif ligand 2, a member of the GPCR family, enhanced somatic cell reprogramming 12.3 times. Furthermore, data from the analysis of the signature gene expression profiles of mouse embryonic fibroblasts at 2 days after RTF infection revealed that the genetic modifier, Whsc1l1 (variant 1), also improved the efficiency of somatic cell reprogramming. Finally, comparison of the overall gene expression profiles between the high and low efficiency conditions will provide novel insights into mechanisms underlying somatic cell reprogramming.
    Reprogramming
    KLF4
    Homeobox protein NANOG
    Citations (33)
    Metadichol, a natural ligand for the expression of Yamanaka reprogramming factors in somatic and primary cancer cell lines
    Research Square (Research Square) (2022)
    P.R. Raghavan
    Abstract The conversion of somatic cells back into induced pluripotent stem cells (iPSCs) or embryonic-like stem cells involves the introduction of four genes commonly called Yamanaka factors: SOX2, OCT4, KLF4, and c-MYC. One drawback is that these genes and the viral vectors used to introduce them into cells have the potential to cause cancer, and these iPSC lines are not clinically useful. Most instances of direct reprogramming have been achieved by the forced expression of defined factors using viral vectors. In this paper, we show that Metadichol® has the potential to generate iPSCs non-virally and may be helpful in clinical applications. Metadichol is a Nano formulation of long-chain alcohols derived from food. Quantitative real-time PCR (qRT-PCR) and western blotting showed that OCT4, SOX2, and NANOG are expressed when fibroblasts were treated with Metadichol at concentrations of one picogram to 100 nanograms. Reverse-transcription PCR (RT-PCR) resulted in OCT4, KLF4, NANOG, and SOX2 levels increased relative to controls by 4.01, 3.51 and 1.26, and 2.5 times, respectively. Such multifold increases were also seen in A549 cancer cell lines and in Colo-205 cells. Reprogramming human primary cancer cells remains a challenge. We present data that even primary cancer cells show increases in the four Yamanaka factors. Metadichol treatment of triple-negative primary breast cancer (HCAF-TNPBC) primary cancer cells led to increases in OKSM factors by 19.6, 8.07, 2.45, and 6.91 times in concentration ranges of 1 picogram to 100 nanograms. Metadichol induces the expression of Klotho, an antiaging gene, that curbs the expression of the TP53 gene, as this is a pre-requisite for reprogramming somatic cells into iPSCs. Metadichol also increases endogenous vitamin C levels, which is necessary for the efficient reprogramming of somatic cells into iPSCs. Metadichol an extract from food that is nontoxic, which opens the door to direct testing in humans with the potential for applications in stem cell therapy
    Reprogramming
    Cancer cell lines
    Expression (computer science)
    Citations (9)
    Optimal reprogramming factor stoichiometry increases colony numbers and affects molecular characteristics of murine induced pluripotent stem cells
    Cytometry Part A (2011)
    Ulf TiemannMalte SgoddaEva WarlichMatthias BallmaierHans R. Schöler
    Abstract Somatic cells can be reprogrammed toward pluripotency by overexpression of a set of transcription factors, yielding induced pluripotent stem cells (iPSCs) with features similar to embryonic stem cells. Little is known to date about stoichiometric requirements of the individual reprogramming factors (RFs) for efficient reprogramming and especially about whether stoichiometry also influences the quality of derived iPSCs. To address this important issue, we chose bicistronic lentiviral vectors coexpressing fluorescent reporters (eGFP, dTomato, Cerulean, or Venus) along with the canonical RFs to transduce a bulk of murine embryonic fibroblasts (MEFs). Using a flow cytometric approach, we were able to independently and proportionally quantify all fluorophores in multiple‐infected MEFs and more importantly could sort these cells into all 16 stoichiometric combinations of high or moderate expression of the four factors. On average, we obtained about 600 alkaline phosphatase‐expressing colonies from 20,000 seeded cells. Interestingly, only seven different stoichiometric ratios gave rise to any colonies at all. The by far most colonies were obtained from those fractions, where Oct4 was in excess over the other three factors (2,386 colonies/20,000 cells), or where both Oct4 and c‐Myc were in excess over Sox2 and Klf4 (1,593 colonies/20,000 cells). Our findings suggest that increased Oct4 levels opposite to modest ones for Sox2 and Klf4 are required for satisfying reprogramming efficiencies and that these stoichiometries are also highly beneficial for achieving a stable pluripotent state independent of ectopic RF expression. Finally, the eligible Oct4 high , Sox2 low , and Klf4 low subpopulation only resembles a small fraction of cells targeted by equal vector amounts, suggesting the necessity to address stoichiometry also in alternative approaches for iPSC generation or between different experimental systems. © 2011 International Society for Advancement of Cytometry.
    KLF4
    Reprogramming
    Citations (72)