Oct4 Expression and Mesenchymal Stromal Cell Plasticity

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2899-2899
Author(s):  
Gustavo Yannarelli ◽  
Huijie Jiang ◽  
Sonia Montanari ◽  
Simone Helke ◽  
Xinghua Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Transcription Factors ◽  
Embryonic Stem ◽  
Mrna Levels ◽  
Oct4 Expression ◽  
Culture Cells ◽  
Oct4 Protein ◽  
Embryonic Cardiomyocytes ◽  
Induced Pluripotent

Abstract Mesenchymal stromal cells (MSCs) have the capacity to differentiate along multiple lineages and are now in cell therapy clinical trials, especially for injured myocardium. Mechanisms mediating tissue regeneration remain unclear and are likely multifactorial. We recently showed that bone marrow-derived murine MSCs can acquire cardiac markers but retain MSCs properties when co-cultured with rat embryonic cardiomyocytes (RECs) (Rose et. al., Stem Cells. Aug7, 2008). The aim of our study was to determine whether expression of the embryonic transcription factor, Oct4 was modulated in this model of MSC plasticity. Wild-type Fvb mouse MSCs (passage 4) were co-cultured with RECs for 5 days and expression of Oct4, Nanog and Sox2, was analyzed by qRT-PCR with mouse-specific primers. Oct4 protein was assessed by immunocytochemistry (ICC) and flow cytometry (FC). The MSCs expressed Oct4, Nanog, and Sox2 transcripts. After co-culture, mRNA levels for Oct4 and Sox2 were upregulated 2.6±1.2-fold (p<0.05) and 2.4±0.4-fold (p<0.05), respectively, compared with MSCs controls, in contrast to Nanog gene expression which remained unchanged (1.2±0.3-fold; p=ns). 83±9% of MSCs nuclei were positive for Oct4 by ICC. To distinguish mouse from rat cells in co-culture, cells were stained with an anti-mouse CD44 antibody which does not cross react with rat CD44. CD44 is expressed on all MSCs and absent on RECs. Flow cytometry showed that Oct4 was over-expressed in CD44+ cells after co-culture. Our data demonstrate that these embryonic transcription factors are constitutively expressed in murine MSCs and that Oct4 and Sox2 transcript levels are increased after co-culture with RECs. Oct4, Nanog and Sox2 are known to maintain pluripotency of embryonic stem cells. Moreover, induced pluripotent stem cells can be generated from mouse fibroblasts by the introduction of Oct4 and Sox2 without the need for Nanog (Cell126:663,2006). Our data infer that embryonic transcription factors are involved in MSCs progression to different cell lineages. Our findings suggest new mechanisms that may mediate MSC plasticity.

scholarly journals Biological importance of OCT transcription factors in reprogramming and development

2021 ◽  
Author(s):  
Kee-Pyo Kim ◽  
Dong Wook Han ◽  
Johnny Kim ◽  
Hans R. Schöler
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Donor Cell ◽  
Structural Components ◽  
Oct4 Expression ◽  
Reprogramming Factors ◽  
Induced Pluripotent

AbstractEctopic expression of Oct4, Sox2, Klf4 and c-Myc can reprogram somatic cells into induced pluripotent stem cells (iPSCs). Attempts to identify genes or chemicals that can functionally replace each of these four reprogramming factors have revealed that exogenous Oct4 is not necessary for reprogramming under certain conditions or in the presence of alternative factors that can regulate endogenous Oct4 expression. For example, polycistronic expression of Sox2, Klf4 and c-Myc can elicit reprogramming by activating endogenous Oct4 expression indirectly. Experiments in which the reprogramming competence of all other Oct family members tested and also in different species have led to the decisive conclusion that Oct proteins display different reprogramming competences and species-dependent reprogramming activity despite their profound sequence conservation. We discuss the roles of the structural components of Oct proteins in reprogramming and how donor cell epigenomes endow Oct proteins with different reprogramming competences.

scholarly journals A mini review on production of pluripotency factors (Oct4, Sox2, Klf4 and c-Myc) through recombinant protein technology

2020 ◽  
Vol 5 (1) ◽  
pp. 1-4 ◽  
Author(s):  
David Septian Sumanto Marpaung ◽  
Ayu Oshin Yap Sinaga
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Embryonic Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Cell Types ◽  
Induced Pluripotency ◽  
Pluripotency Factors ◽  
Induced Pluripotent

The four transcription factors OCT4, SOX2, KLF4 and c-MYC are highly expressed in embryonic stem cells (ESC) and their overexpression can induce pluripotency, the ability to differentiate into all cell types of an organism. The ectopic expression such transcription factors could reprogram somatic stem cells become induced pluripotency stem cells (iPSC), an embryonic stem cells-like. Production of recombinant pluripotency factors gain interests due to high demand from generation of induced pluripotent stem cells in regenerative medical therapy recently. This review will focus on demonstrate the recent advances in recombinant pluripotency factor production using various host.

scholarly journals Generation of induced pluripotent stem cells from human blood

Blood ◽  
2009 ◽  
Vol 113 (22) ◽  
pp. 5476-5479 ◽  
Author(s):  
Yuin-Han Loh ◽  
Suneet Agarwal ◽  
In-Hyun Park ◽  
Achia Urbach ◽  
Hongguang Huo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Human Blood ◽  
Pluripotent Stem Cells ◽  
Methylation Status ◽  
Embryonic Stem ◽  
Dermal Fibroblasts ◽  
Patient Specific ◽  
Induced Pluripotent

Human dermal fibroblasts obtained by skin biopsy can be reprogrammed directly to pluripotency by the ectopic expression of defined transcription factors. Here, we describe the derivation of induced pluripotent stem cells from CD34+ mobilized human peripheral blood cells using retroviral transduction of OCT4/SOX2/KLF4/MYC. Blood-derived human induced pluripotent stem cells are indistinguishable from human embryonic stem cells with respect to morphology, expression of surface antigens, and pluripotency-associated transcription factors, DNA methylation status at pluripotent cell-specific genes, and the capacity to differentiate in vitro and in teratomas. The ability to reprogram cells from human blood will allow the generation of patient-specific stem cells for diseases in which the disease-causing somatic mutations are restricted to cells of the hematopoietic lineage.

scholarly journals Inducible expression of Oct-3/4 reveals synergy with Klf4 in targeting Cyclin A2 to enhance proliferation during early reprogramming

2021 ◽  
Author(s):  
Lamuk Zaveri ◽  
Jyotsna Dhawan
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Embryonic Stem ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Mesenchymal To Epithelial Transition ◽  
Cyclin A2 ◽  
Slow Cycling ◽  
Induced Pluripotent ◽  
Dose Dependent

AbstractDuring reprogramming of somatic cells, heightened proliferation is one of the earliest changes observed. While other early events such as mesenchymal-to-epithelial transition have been well studied, the mechanisms by which the cell cycle switches from a slow cycling state to a faster cycling state are still incompletely understood. To investigate the role of Oct-3/4 in this early feature of reprogramming, we created a 4-Hydroxytamoxifen dependent Oct-3/4 Estrogen Receptor fusion (OctER). We show that OctER can substitute for Oct-3/4 to reprogram mouse embryonic fibroblasts to induced pluripotent stem cells. While over-expression of OctER or Klf4 individually did not affect cell proliferation, in combination, these factors hasten the cell cycle, in a tamoxifen dose-dependent manner, supporting a key role for OctER. Oct-3/4 + Klf4 increased proliferation by enhancing expression of Cyclin A2. We verified occupancy of endogenous Oct-3/4 and Klf4 at bioinformatically identified binding sites in the Cyclin A2 promoter in mouse embryonic stem cells (mESC). Using inducible OctER along with Klf4, we show dose-dependent induction of Cyclin A2 promoter-reporter activity and mRNA levels. Taken together, our results provide further evidence of the interdependence of pluripotency and the rapid cell cycle seen in mESC, and identify CyclinA2 as a key early target.

scholarly journals La reprogramación en la obtención de células madre pluripotentes inducidas

2018 ◽  
pp. 9-14
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Phenotypic Characteristics ◽  
Ethical Problems ◽  
Apartado Postal ◽  
Induced Pluripotent ◽  
Factor C

La reprogramación en la obtención de células madre pluripotentes inducidas Reprogramming in obtaining induced pluripotent stem cells   Luis Fernando Tume Farfán  Universidad Nacional de Piura, Urb. Miraflores s/n, Castilla – Apartado Postal 295, Piura, Perú DOI: https://doi.org/10.33017/RevECIPeru2013.0002/ Resumen Debido a los problemas éticos que se han venido dando por el uso de embriones como fuente de células pluripotentes, se han desarrollado nuevas fuentes para obtener células con las mismas características. En esta revisión se discuten algunos métodos de reprogramación que se han sido empleados por muchos investigadores alrededor del mundo, partiendo por la trasferencia nuclear y posteriormente con el trabajo de Yamanaka quien empezó a usar la introducción de los cuatro factores de transcripción, Oct3 / 4, Sox2, Klf4 y c-Myc que origina con éxito la reprogramación de las células somáticas en células madre pluripotentes inducidas (iPSC), que poseen características genómicas y fenotípicas de células madre embrionarias, además se describe algunas desventajas que tienen estos métodos y los riesgos que involucran el uso del factor de transcripción c-Myc. A pesar de que queda mucho por mejorar en este campo, las células iPS muestran un tremendo potencial para la investigación y sus posibles aplicaciones terapéuticas en la medicina regenerativa. Descriptores: Pluripotenciales, factores de transcripción, inducidas, reprogramación. Abstract Because of the ethical problems that have been taking for the use of embryos as a source of pluripotent cells , we have developed new sources for cells with the same characteristics. In this review we discuss some reprogramming methods that have been employed by many researchers around the world , starting with the nuclear transfer and later with the work of Yamanaka who began using the introduction of four transcription factors , Oct3 / 4 , Sox2 , Klf4 and c- Myc that originates successful reprogramming of somatic cells into induced pluripotent stem cells ( iPSC ) , which have genomic and phenotypic characteristics of embryonic stem cells , and discusses some drawbacks with these methods and the risks involve the use of the transcription factor c -Myc . Although much room for improvement in this field, iPS cells show a tremendous potential for research and potential therapeutic applications in regenerative medicine. Keywords: Keywords: Pluripotent transcription factors, induced, reprogramming.

scholarly journals Ten years of progress and promise of induced pluripotent stem cells: historical origins, characteristics, mechanisms, limitations, and potential applications

2017 ◽  
Author(s):  
Adekunle Ebenezer Omole ◽  
Adegbenro Omotuyi John Fakoya
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Molecular Mechanism ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Patient Specific ◽  
Induced Pluripotent

The discovery of induced pluripotent stem cells (iPSCs) by Shinya Yamanaka in 2006 was heralded as a major breakthrough of the decade in stem cell research. The ability to reprogrammed human somatic cells to a pluripotent embryonic stem cell-like state through the ectopic expression of a combination of embryonic transcription factors was greeted with great excitement by scientists and bioethicists. The reprogramming technology offers the opportunity to generate patient-specific stem cells for modeling human diseases, drug development and screening, and individualized regenerative cell therapy. However, fundamental questions have been raised regarding the molecular mechanism of iPSCs generation, a process still poorly understood by scientists. The efficiency of reprogramming of iPSCs remains low due to the effect of various barriers of reprogramming. There is also the risk of chromosomal instability and oncogenic transformation associated with the use of viral vectors, such as retrovirus and lentivirus, which deliver the reprogramming transcription factors by integration in the host cell genome. These challenges can hinder the therapeutic prospects and promise of iPSCs and their clinical applications. Consequently, extensive studies have been done to elucidate the molecular mechanism of reprogramming and novel strategies have been identified which help to improve the efficiency of reprogramming methods and overcome the safety concerns linked with iPSCs generation. Distinct barriers and enhancers of reprogramming have been elucidated and non-integrating reprogramming methods have been reported. Here, we summarize the progress and the recent advances that have been made over the last 10 years in the iPSCs field, with emphasis on the molecular mechanism of reprogramming, strategies to improve the efficiency of reprogramming, characteristics and limitations of iPSCs, and the progress made in the applications of iPSCs in the field of disease modelling, drug discovery and regenerative medicine. Additionally, this study appraised the role of genomic editing technology in the generation of healthy iPSCs.

scholarly journals Effect of exogenous transcription factors integration sites on safety and pluripotency of induced pluripotent stem cells

2020 ◽  
Vol 23 (1) ◽  
pp. 5-13
Author(s):  
S Yin ◽  
W Li ◽  
G Yang ◽  
Y Cheng ◽  
Q Yi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Ontology ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Tetraploid Complementation ◽  
Integration Sites ◽  
Exogenous Genes ◽  
Induced Pluripotent

AbstractInduced pluripotent stem cells (iPSCs), generated from somatic cells, not only possess similar characteristics with embryonic stem cells (ESCs), but also present more advantages than ESCs in medical applications. The classical induction method that utilizes the integration of exogenous genes into chromosomes may raise the potential risk of the safety of iPSCs. To investigate the potential correlation between the integration sites of exogenous transcription factors (TFs) and iPSCs’ pluripotency and safety, the integration of exogenous genes in three iPSC lines, which met the golden standard of murine developmental assay (tetraploid complementation), were analyzed. Twenty-two integration sites of exogenous TFs were identified by nested inverse polymerase chain reaction (iPCR) and 39 flanking genes’ functions were analyzed by gene ontology (GO). In the 22 integrated sites, 17 (77.3%) were located in the intergenic regions and the remainder were located in introns far from the transcription start sites. Microarray analysis of the flanking genes in these cells showed that there was no distinct difference in expression levels between the iPSCs, ESCs and mouse embryonic fibroblast (MEF), suggesting that the integration of exogenous TFs has no significant influence on the expression of flanking genes. Gene ontology analysis showed that although most of the flanking genes were housekeeping genes, which were necessary for basic life activity, none of these 39 flanking genes have correlation with tumorigenesis or embryogenesis, suggesting that the integration sites hold low risk of tumorigenesis.

scholarly journals Ten years of progress and promise of induced pluripotent stem cells: historical origins, characteristics, mechanisms, limitations, and potential applications

2017 ◽  
Author(s):  
Adekunle Ebenezer Omole ◽  
Adegbenro Omotuyi John Fakoya
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Molecular Mechanism ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Patient Specific ◽  
Induced Pluripotent

The discovery of induced pluripotent stem cells (iPSCs) by Shinya Yamanaka in 2006 was heralded as a major breakthrough of the decade in stem cell research. The ability to reprogrammed human somatic cells to a pluripotent embryonic stem cell-like state through the ectopic expression of a combination of embryonic transcription factors was greeted with great excitement by scientists and bioethicists. The reprogramming technology offers the opportunity to generate patient-specific stem cells for modeling human diseases, drug development and screening, and individualized regenerative cell therapy. However, fundamental questions have been raised regarding the molecular mechanism of iPSCs generation, a process still poorly understood by scientists. The efficiency of reprogramming of iPSCs remains low due to the effect of various barriers of reprogramming. There is also the risk of chromosomal instability and oncogenic transformation associated with the use of viral vectors, such as retrovirus and lentivirus, which deliver the reprogramming transcription factors by integration in the host cell genome. These challenges can hinder the therapeutic prospects and promise of iPSCs and their clinical applications. Consequently, extensive studies have been done to elucidate the molecular mechanism of reprogramming and novel strategies have been identified which help to improve the efficiency of reprogramming methods and overcome the safety concerns linked with iPSCs generation. Distinct barriers and enhancers of reprogramming have been elucidated and non-integrating reprogramming methods have been reported. Here, we summarize the progress and the recent advances that have been made over the last 10 years in the iPSCs field, with emphasis on the molecular mechanism of reprogramming, strategies to improve the efficiency of reprogramming, characteristics and limitations of iPSCs, and the progress made in the applications of iPSCs in the field of disease modelling, drug discovery and regenerative medicine. Additionally, this study appraised the role of genomic editing technology in the generation of healthy iPSCs.

scholarly journals Ten years of progress and promise of induced pluripotent stem cells: historical origins, characteristics, mechanisms, limitations, and potential applications

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4370 ◽  
Author(s):  
Adekunle Ebenezer Omole ◽  
Adegbenro Omotuyi John Fakoya
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Molecular Mechanism ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Patient Specific ◽  
Induced Pluripotent

The discovery of induced pluripotent stem cells (iPSCs) by Shinya Yamanaka in 2006 was heralded as a major breakthrough of the decade in stem cell research. The ability to reprogram human somatic cells to a pluripotent embryonic stem cell-like state through the ectopic expression of a combination of embryonic transcription factors was greeted with great excitement by scientists and bioethicists. The reprogramming technology offers the opportunity to generate patient-specific stem cells for modeling human diseases, drug development and screening, and individualized regenerative cell therapy. However, fundamental questions have been raised regarding the molecular mechanism of iPSCs generation, a process still poorly understood by scientists. The efficiency of reprogramming of iPSCs remains low due to the effect of various barriers to reprogramming. There is also the risk of chromosomal instability and oncogenic transformation associated with the use of viral vectors, such as retrovirus and lentivirus, which deliver the reprogramming transcription factors by integration in the host cell genome. These challenges can hinder the therapeutic prospects and promise of iPSCs and their clinical applications. Consequently, extensive studies have been done to elucidate the molecular mechanism of reprogramming and novel strategies have been identified which help to improve the efficiency of reprogramming methods and overcome the safety concerns linked with iPSC generation. Distinct barriers and enhancers of reprogramming have been elucidated, and non-integrating reprogramming methods have been reported. Here, we summarize the progress and the recent advances that have been made over the last 10 years in the iPSC field, with emphasis on the molecular mechanism of reprogramming, strategies to improve the efficiency of reprogramming, characteristics and limitations of iPSCs, and the progress made in the applications of iPSCs in the field of disease modelling, drug discovery and regenerative medicine. Additionally, this study appraises the role of genomic editing technology in the generation of healthy iPSCs.

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