Nuclear reprogramming and induced pluripotency

2019 ◽  
pp. 205-212
Author(s):  
John C. Lucchesi
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Late Onset ◽  
Neurological Diseases ◽  
Embryonic Stem ◽  
Induced Pluripotency ◽  
Differentiated Cells ◽  
Induced Pluripotent ◽  
Bivalent Promoter

Four core transcription factors known to maintain the pluripotent state in embryonic stem cells (ESCs)—Oct4, Sox2, Klf4 and c-Myc—were used to induce pluripotent stem cells in adult-derived fibroblasts. Induced pluripotent stem cells (iPSCs), like ESCs, have less condensed and more transcriptionally active chromatin than differentiated cells. The number of genes with bivalent promoter marks increases during reprogramming, reflecting the switch of differentiation-specific active genes to an inactive, but poised, status. The levels of DNA methyl transferases and demethylases are increased, underlying the changes in the pattern of DNA methylation that occur late during reprogramming. The potential therapeutic applications of iPSCs include reprogramming a patient’s own cells to avoid the problem of rejection following injection to restore tissue or organ function. iPSCs derived from individuals at risk of developing late-onset neurological diseases could be differentiated in culture to predict the future occurrence of the disease. Caveats involve the fact that long-term culturing often results in genomic mutations that may, by chance, involve tumor suppressors or oncogenes.

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 Identification of Molecular Signatures in Neural Differentiation and Neurological Diseases Using Digital Color-Coded Molecular Barcoding

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Debora Salerno ◽  
Alessandro Rosa
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Neural Differentiation ◽  
Disease Modeling ◽  
Neurological Diseases ◽  
Embryonic Stem ◽  
Molecular Signatures ◽  
Molecular Barcoding ◽  
Induced Pluripotent

Human pluripotent stem cells (PSCs), including embryonic stem cells and induced pluripotent stem cells, represent powerful tools for disease modeling and for therapeutic applications. PSCs are particularly useful for the study of development and diseases of the nervous system. However, generating in vitro models that recapitulate the architecture and the full variety of subtypes of cells that make the complexity of our brain remains a challenge. In order to fully exploit the potential of PSCs, advanced methods that facilitate the identification of molecular signatures in neural differentiation and neurological diseases are highly demanded. Here, we review the literature on the development and application of digital color-coded molecular barcoding as a potential tool for standardizing PSC research and applications in neuroscience. We will also describe relevant examples of the use of this technique for the characterization of the heterogeneous composition of the brain tumor glioblastoma multiforme.

scholarly journals Mammalian genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and invertebrate species

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
Ricardo Antonio Rosselló ◽  
Chun-Chun Chen ◽  
Rui Dai ◽  
Jason T Howard ◽  
Ute Hochgeschwender ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cells ◽  
Mammalian Species ◽  
Embryonic Stem ◽  
Model Organisms ◽  
Differentiated Cells ◽  
Transcription Factor Genes ◽  
Induced Pluripotent

Cells are fundamental units of life, but little is known about evolution of cell states. Induced pluripotent stem cells (iPSCs) are once differentiated cells that have been re-programmed to an embryonic stem cell-like state, providing a powerful platform for biology and medicine. However, they have been limited to a few mammalian species. Here we found that a set of four mammalian transcription factor genes used to generate iPSCs in mouse and humans can induce a partially reprogrammed pluripotent stem cell (PRPSCs) state in vertebrate and invertebrate model organisms, in mammals, birds, fish, and fly, which span 550 million years from a common ancestor. These findings are one of the first to show cross-lineage stem cell-like induction, and to generate pluripotent-like cells for several of these species with in vivo chimeras. We suggest that the stem-cell state may be highly conserved across a wide phylogenetic range.

Differentiation of human induced pluripotent stem cells into testosterone-producing Leydig-like cells

Endocrinology ◽  
2021 ◽  
Author(s):  
Takaki Ishida ◽  
Michiyo Koyanagi-Aoi ◽  
Daisuke Yamamiya ◽  
Atsushi Onishi ◽  
Katsuya Sato ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Leydig Cells ◽  
Late Onset ◽  
Testosterone Replacement Therapy ◽  
Promising Alternative ◽  
Differentiated Cells ◽  
Induced Pluripotent ◽  
Late Onset Hypogonadism

Abstract Late-onset hypogonadism (LOH) syndrome due to a partial lack of testosterone, which is mainly secreted by Leydig cells in the testes, decreases the quality of life of older men. Leydig cell transplantation is expected to be a promising alternative to conventional testosterone replacement therapy (TRT) for LOH syndrome. We herein report a simple and robust protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into Leydig-like cells by doxycycline-inducible overexpression of NR5A1 and treatment with a combination of 8-bromoadenosine-3’,5’-cyclic monophosphate (8-Br-cAMP) and forskolin. The differentiated cells expressed the steroidogenic enzyme genes STAR, CYP11A1, CYP17A1 and HSD3B2 and the specific markers of adult Leydig cells HSD17B3, INSL3 and LHCGR. Furthermore, we confirmed the secretion of functional testosterone from the cells into the culture supernatant by a testosterone-sensitive cell proliferation assay. These findings showed that the hiPSCs were able to be differentiated into Leydig-like cells, supporting the expectation that hiPSC-derived Leydig-like cells can be novel tools for treating LOH syndrome.

scholarly journals The genetics of induced pluripotency

Reproduction ◽  
2010 ◽  
Vol 139 (1) ◽  
pp. 35-44 ◽  
Author(s):  
Amy Ralston ◽  
Janet Rossant
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Molecular Level ◽  
Embryonic Stem ◽  
Cell Types ◽  
Therapeutic Benefit ◽  
Mature Cell ◽  
Developmental Potential ◽  
Induced Pluripotency ◽  
Induced Pluripotent

The flurry of recent publications regarding reprogramming of mature cell types to induced pluripotent stem cells raises the question: what exactly is pluripotency? A functional definition is provided by examination of the developmental potential of pluripotent stem cell types. Defining pluripotency at the molecular level, however, can be a greater challenge. Here, we examine the emerging list of genes associated with induced pluripotency, with particular attention to their functional requirement in the mouse embryo. Knowledge of the requirement for these genes in the embryo and in embryonic stem cells will advance our understanding of how to reverse the developmental clock for therapeutic benefit.

IDENTIFICATION of Infrared SPECTRAL Signature of INDUCED Pluripotent STEM CELLS (iPSC)

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4792-4792
Author(s):  
Christophe Sandt ◽  
Olivier Feraud ◽  
Ali G. Turhan ◽  
Paul Dumas ◽  
Annelise Bennaceur-Griscelli
Keyword(s):  
Stem Cells ◽  
Cell Line ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Spectral Signature ◽  
Es Cell ◽  
Differentiated Cells ◽  
Murine Fibroblasts ◽  
Infrared Spectral ◽  
Induced Pluripotent

Abstract Abstract 4792 Recent technological advances in cell reprogramming by generation of induced pluripotent stem cells (iPSC) offer major perspectives in disease modelling and future hopes for providing novel stem cells sources in regenerative medicine. However, research on iPSC still requires refining the criteria of the pluripotency stage of these cells and exploration of their equivalent functionality to human embryonic stem cells (hESC). In this work, we report that the use of the Synchrotron-based FTIR microspectroscopy allows following the infrared spectral modification of the differentiated cells during the reprogramming process as well as the comparison between iPSC with hESC. The model that we studied consisted on the use of human ES cell line H9 grown on murine embryonic fibroblasts in the presence of bFGF. We have generated mesenchymal stem cells from the H9 cell line (H9-MSC) and we used them to generate iPSC (iPSC-H9) by the enforced expression of pluripotency genes Oct4, Sox2, Lin28 and Nanog. We have also followed the same approach on murine cells by generating murine iPSC from murine ES cells by retrovirus mediated gene transfer of Oct4, Sox2, c-Myc and Klf4. iPSC were characterized by expression of pluripotency markers, and teratoma assays. Infrared Spectral fingerprints of the original H9, MSC-H9 and iPSC-H9 as well as differentiated murine fibroblasts and murine iPSC were acquired at sub-cellular resolution using a synchrotron-powered infrared microscope. In murine system, the spectral signature of iPSC has been compared to that of D3, a well-characterized murine ES cell line. The spectral signature of iPSC and ESC displays a marked difference with those of the differentiated cells used before reprogrammation regardless the origin of the target cell (mesenchymal stem cells or murine fibroblasts). We unambiguously demonstrate for the first time to our knowledge, that the human and murine iPSC retrieve the same chemical composition with an indistinguishable spectral signature from their embryonic stem cells counterparts. Importantly, the spectral signatures were found to be specific to each of the cell line, as evidenced using pattern recognition methods and illustrated the genetic biodiversity of each iPSC and ESC. Thus, in addition to the classical pluripotency markers, FTIR microspectroscopy signature could be a rapid methodology to evaluate the pluripotency after somatic cell reprogramming. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Integrative Research of Induction of Pluripotent Stem Cells

2017 ◽  
Vol 4 (3-4) ◽  
pp. 115-124 ◽  
Author(s):  
Penglin Gao ◽  
Chuanhe Sun ◽  
Weilong Liao ◽  
Wenfei Jiang ◽  
Te Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Viral Vector ◽  
Neurological Diseases ◽  
Embryonic Stem ◽  
Basic Research ◽  
Model Systems ◽  
Suitable Model ◽  
Integrative Research ◽  
Induced Pluripotent

Since the Japanese scientist Shinya Yamanaka used a viral vector to transfer the combination of 4 factors into differentiated somatic cells and reprogramed them to obtain similar embryonic stem cells and induced pluripotent stem cells (iPSCs), it provided one integrative method for studying many medical fields. Patient-derived iPSCs have provided an opportunity to study human diseases for which no suitable model systems are available. iPSC technology has since become a major breakthrough in the field of stem cell research. With the continuous development of iPSC technology and the continuous improvement of technical levels, excellent advances have become more and more common in the basic research and medical fields of life sciences. This article reviews the development history, clinical application, and problems and prospects of iPSC, and focuses on the application of iPSC in neurological diseases.

scholarly journals Adhesive Signature-Based, Label-Free Isolation of Human Pluripotent Stem Cells

2012 ◽  
Author(s):  
Ankur Singh ◽  
Shalu Suri ◽  
Ted T. Lee ◽  
Jamie M. Chilton ◽  
Steve L. Stice ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Somatic Cells ◽  
Embryonic Stem ◽  
Mouse Embryonic Fibroblast ◽  
Feeder Layer ◽  
Label Free ◽  
Differentiated Cells ◽  
Cell Sources ◽  
Induced Pluripotent

Generation of human induced pluripotent stem cells (hiPSCs) from fibroblasts and other somatic cells represents a highly promising strategy to produce auto- and allo-genic cell sources for therapeutic approaches as well as novel models of human development and disease1. Reprogramming protocols involve transduction of the Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc into the parental somatic cells, followed by culturing the transduced cells on mouse embryonic fibroblast (MEF) or human fibroblast feeder layers, and subsequent mechanical dissociation of pluripotent cell-like colonies for propagation on feeder layers1, 2. The presence of residual parental and feeder-layer cells introduces experimental variability, pathogenic contamination, and promotes immunogenicity3. Similar to human embryonic stem cells (hESCs), reprogrammed hiPSCs suffer from the unavoidable problem of spontaneous differentiation due to sub-optimal feeder cultures4, growth factors5, and the feeder-free substrate6. Spontaneously differentiated (SD)-hiPSCs display reduced pluripotency and often contaminate hiPSC cultures, resulting in overgrowth of cultures and compromising the quality of residual pluripotent stem cells5. Therefore, the ability to rapidly and efficiently isolate undifferentiated hiPSCs from the parental somatic cells, feeder-layer cells, and spontaneously differentiated cells is a crucial step that remains a bottleneck in all human pluripotent stem cell research.

scholarly journals Maintenance and Neuronal Differentiation of Chicken Induced Pluripotent Stem-Like Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-14 ◽  
Author(s):  
Rui Dai ◽  
Ricardo Rossello ◽  
Chun-chun Chen ◽  
Joeran Kessler ◽  
Ian Davison ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neuronal Differentiation ◽  
Pluripotent Stem Cells ◽  
Vocal Learning ◽  
Culture Conditions ◽  
Differentiated Cells ◽  
Induced Pluripotent ◽  
Stem Cell State

Pluripotent stem cells have the potential to become any cell in the adult body, including neurons and glia. Avian stem cells could be used to study questions, like vocal learning, that would be difficult to examine with traditional mouse models. Induced pluripotent stem cells (iPSCs) are differentiated cells that have been reprogrammed to a pluripotent stem cell state, usually using inducing genes or other molecules. We recently succeeded in generating avian iPSC-like cells using mammalian genes, overcoming a limitation in the generation and use of iPSCs in nonmammalian species (Rosselló et al., 2013). However, there were no established optimal cell culture conditions for avian iPSCs to establish long-term cell lines and thus to study neuronal differentiationin vitro. Here we present an efficient method of maintaining chicken iPSC-like cells and for differentiating them into action potential generating neurons.

scholarly journals Ammonia-based enrichment and long-term propagation of zone I hepatocyte-like cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ruri Tsuneishi ◽  
Noriaki Saku ◽  
Shoko Miyata ◽  
Saeko Akiyama ◽  
Palaksha Kanive Javaregowda ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Selection Strategy ◽  
Ammonia Metabolism ◽  
Cytochrome P450 Genes ◽  
Induced Pluripotent ◽  
Selection Agent

AbstractAmmonia has a cytotoxic effect and can therefore be used as a selection agent for enrichment of zone I hepatocytes. However, it has not yet been determined whether ammonia-treated hepatocyte-like cells are able to proliferate in vitro. In this study, we employed an ammonia selection strategy to purify hepatocyte-like cells that were differentiated from human embryonic stem cells (ESCs) and from induced pluripotent stem cells (iPSCs). The resistance to cytotoxicity or cell death by ammonia is likely attributable to the metabolism of ammonia in the cells. In addition to ammonia metabolism-related genes, ammonia-selected hepatocytes showed increased expression of the cytochrome P450 genes. Additionally, the ammonia-selected cells achieved immortality or at least an equivalent life span to human pluripotent stem cells without affecting expression of the liver-associated genes. Ammonia treatment in combination with in vitro propagation is useful for obtaining large quantities of hepatocytes.

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