The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform

Junghyun Kim - One of the best experts on this subject based on the ideXlab platform.

  • establishment of a tlr3 homozygous knockout human induced pluripotent Stem Cell Line using crispr cas9
    Stem Cell Research, 2021
    Co-Authors: Junghyun Kim, Hyeongjun Han
    Abstract:

    Abstract The Toll-like receptor (TLR) family plays an important role in the recognition of pathogens, including bacteria, viruses, fungi, and parasites, followed by the activation of innate immunity. TLR3 recognizes double-stranded RNA, a form of genetic material produced by positive-strand RNA viruses and DNA viruses, and is activated by viral infection. Upon recognition, TLR3 promotes the activation of interferon regulatory factor 3 to enhance the expression and secretion of type I interferons that signal other Cells to enhance their antiviral defenses. We generated biallelic mutants of the TLR3 gene using a CRISPR-Cas9 genome editing method in human induced pluripotent Stem Cells (hiPSCs). TLR3 homozygous-knockout hiPSCs retained normal morphology, gene expression, and in vivo differentiation potential.

  • generation of a nrf2 homozygous knockout human embryonic Stem Cell Line using crispr cas9
    Stem Cell Research, 2017
    Co-Authors: Sojung Kim, Omer Habib, Jinsoo Kim, Hyowon Han, Soo Kyung Koo, Junghyun Kim
    Abstract:

    Nuclear factor erythroid 2-related factor 2 (NFE2L2 or Nrf2) is a well-known transcription factor that regulates the expression of a large number of anti-oxidant genes in mammalian Cells (J.H. Kim et al., 2014). Here, we generated a homozygous Nrf2 knockout human embryonic Stem Cell (hESC) Line, H9Nrf2KO-A13, using the CRISPR/Cas9 genome editing method. The Nrf2 homozygous knockout H9 Cell Line maintains pluripotency, differentiation potential into three germ layers, and a normal karyotype.

  • a homozygous keap1 knockout human embryonic Stem Cell Line generated using crispr cas9 mediates gene targeting
    Stem Cell Research, 2017
    Co-Authors: Sojung Kim, Omer Habib, Jinsoo Kim, Hyowon Han, Soo Kyung Koo, Junghyun Kim
    Abstract:

    Kelch-like ECH-associated protein 1 (keap1) is a cysteine-rich protein that interacts with transcription factor Nrf2 in a redox-sensitive manner, leading to the degradation of Nrf2 (Kim et al., 2014a). Disruption of Keap1 results in the induction of Nrf2-related signaling pathways involving the expression of a set of anti-oxidant and anti-inflammatory genes. We generated biallelic mutants of the Keap1 gene using a CRISPR-Cas9 genome editing method in the H9 human embryonic Stem Cell (hESC). The Keap1 homozygous-knockout H9 Cell Line retained normal morphology, gene expression, and in vivo differentiation potential.

James Adjaye - One of the best experts on this subject based on the ideXlab platform.

  • lymphoblast derived integration free isrm con9 ips Cell Line from a 75year old female
    Stem Cell Research, 2018
    Co-Authors: Soraia Martins, Wasco Wruck, Friederike Schroter, Kristel Sleegers, Martina Bohndorf, Christine Van Broeckhoven, Fatima Assar, James Adjaye
    Abstract:

    Human lymphoblast Cells were used to generate integration-free induced pluripotent Stem Cells (iPSCs) employing episomal-based plasmids expressing OCT4, SOX2, NANOG, LIN28, c-MYC and L-MYC. The derived iPSCs were defined as pluripotent based on (i) expression of pluripotency-associated markers, (ii) embryoid body-based differentiation into Cell types representative of the three germ layers and (iii) the similarity between the transcriptomes of the iPSC Line and the human embryonic Stem Cell Line H1 with a Pearson correlation of 0.95.

  • Generation and characterization of two iPSC Lines from human epicardium-derived Cells
    Elsevier, 2017
    Co-Authors: Christina Paulitschek, Peggy Schulze-matz, Julia Hesse, Timo Schmidt, Wasco Wruck, James Adjaye, Jürgen Schrader
    Abstract:

    Human epicardium-derived Cells (EPDC) were reprogrammed to generate two iPSC Lines, MCDU1i-EPDC and MCDU2i-EPDC, by nucleofection of episomal-based plasmids expressing the reprogramming factors OCT4, SOX2, KLF4, c-MYC, NANOG and LIN28. Pluripotency was confirmed in vitro by immunofluorescence analysis and embryoid body formation. The iPSC Lines and the human embryonic Stem Cell Line H1 show a Pearson correlation co-efficient of 0.951 (MCDU1i-EPDC) and 0.937 (MCDU2i-EPDC) as assessed by comparative transcriptome profiling

  • lymphoblast derived integration free ips Cell Line from a 69 year old male
    Stem Cell Research, 2016
    Co-Authors: Friederike Schroter, Wasco Wruck, Kristel Sleegers, Martina Bohndorf, Christine Van Broeckhoven, James Adjaye
    Abstract:

    Human lymphoblast Cells were used to generate integration-free induced pluripotent Stem (iPS) Cells employing episomal plasmids expressing OCT4, SOX2, NANOG, LIN28, C-MYC and L-MYC. The derived iPS Cells were defined as pluripotent based on (i) expression of pluripotent-associated markers, (ii) embryoid body-based differentiation into Cell types representative of the three germ layers and (iii) the similarity between the transcriptomes of the iPS Cell Line and the human embryonic Stem Cell Line H1 with a Pearson correlation of 0.95.

  • lymphoblast derived integration free ips Cell Line from a 65 year old alzheimer s disease patient expressing the trem2 p r47h variant
    Stem Cell Research, 2016
    Co-Authors: Friederike Schroter, Wasco Wruck, Kristel Sleegers, Martina Bohndorf, Christine Van Broeckhoven, Elise Cuyvers, James Adjaye
    Abstract:

    Human lymphoblast Cells from a male patient diagnosed with Alzheimer's disease (AD) expressing the TREM2 p.R47H variant were used to generate integration-free induced pluripotent Stem (iPS) Cells employing episomal plasmids expressing OCT4, SOX2, NANOG, LIN28, c-MYC and L-MYC. The iPS Cells retained the TREM2 mutation, and were defined as pluripotent based on (i) expression of pluripotent-associated markers, (ii) embryoid body-based differentiation into Cell types representative of the three germ layers and (iii) the similarity between the transcriptomes of the iPS Cell Line and the human embryonic Stem Cell Line H1 with a Pearson correlation of 0.966.

  • generation of ipsc Line epihuvec from human umbilical vein endothelial Cells
    Stem Cell Research, 2015
    Co-Authors: James Adjaye, Peggy Matz
    Abstract:

    Human umbilical vein endothelial Cells (HUVECs) were used to generate the iPSC Line epiHUVEC employing a combination of three episomal-based plasmids expressing OCT4, SOX2, NANOG, LIN28, c-MYC and KLF4. Pluripotency was confirmed both in vivo and in vitro. The transcriptome profile of epiHUVEC and the human embryonic Stem Cell Line - H1 have a Pearson correlation of 0.899.

Henrik Semb - One of the best experts on this subject based on the ideXlab platform.

  • derivation of a xeno free human embryonic Stem Cell Line
    Stem Cells, 2006
    Co-Authors: Catharina Ellerstrom, Raimund Strehl, Karina Moya, Katarina Andersson, Christina Bergh, Kersti Lundin, Johan Hyllner, Henrik Semb
    Abstract:

    Elimination of all animal material during both the derivation and long-term culture of human embryonic Stem Cells (hESCs) is necessary prior to future application of hESCs in clinical Cell therapy. The potential consequences of transplanting xeno-contaminated hESCs into patients, such as an increased risk of graft rejection [Stem CellS 2006;24:221229] and the potential transfer of nonhuman pathogens, make existing hESC Lines unsuitable for clinical applications. To avoid xeno-contamination during derivation and culture of hESCs, we first developed a xeno-free medium supplemented with human serum, which supports long-term (> 50 passages) culture of hESCs in an undifferentiated state. To enable derivation of new xeno-free hESCs, we also established xeno-free human foreskin fibroblast feeders and replaced immunosurgery, which involves the use of guinea pig complement, with a modified animal-product-free derivation procedure. Here, we report the establishment and characterization (> 20 passages) of a xeno-free pluripotent diploid normal hESC Line, SA611.

Lara Stevanato - One of the best experts on this subject based on the ideXlab platform.

  • implantation of the clinical grade human neural Stem Cell Line ctx0e03 rescues the behavioral and pathological deficits in the quinolinic acid lesioned rodent model of huntington s disease
    Stem Cells, 2020
    Co-Authors: Yongwoo Yoon, Lara Stevanato, Iksoo Jeon, Hyun Jung Park, Inhyun Park, Caroline Hicks, Randolph Corteling, Roger A Barker, John Sinden, Jihwan Song
    Abstract:

    : Huntington's disease (HD) is a devastating, autosomal-dominant neurodegenerative disease, for which there are currently no disease-modifying therapies. Clinical trials to replace the damaged striatal medium spiny neurons (MSNs) have been attempted in the past two decades but have met with only limited success. In this study, we investigated whether a clonal, conditionally immortalized neural Stem Cell Line (CTX0E03), which has already shown safety and signals of efficacy in chronic ischemic stroke patients, could rescue deficits seen in an animal model of HD. After CTX0E03 transplantation into the quinolinic acid-lesioned rat model of HD, behavioral changes were measured using the rotarod, stepping, and staircase tests. In vivo differentiation and neuronal connections of the transplanted CTX0E03 Cells were evaluated with immunohistochemical staining and retrograde tracing with Fluoro-Gold. We found that transplantation of CTX0E03 gave rise to a significant behavioral improvement compared with the sham- or fibroblast-transplanted group. Transplanted CTX0E03 formed MSNs (DARPP-32) and GABAergic neurons (GABA, GAD65/67) with BDNF expression in the striatum, while cortically transplanted Cells formed Tbr1-positive neurons. Using a retrograde label, we also found stable engraftment and connection of the transplanted Cells with host brain tissues. CTX0E03 transplantation also reduced glial scar formation and inflammation, as well as increasing endogenous neurogenesis and angiogenesis. Overall, our results demonstrate that CTX0E03, a clinical-grade neural Stem Cell Line, is effective for preclinical test in HD, and, therefore, will be useful for clinical development in the treatment of HD patients.

  • an allogeneic off the shelf therapeutic strategy for peripheral nerve tissue engineering using clinical grade human neural Stem Cells
    Scientific Reports, 2018
    Co-Authors: C Orourke, Lara Stevanato, John Sinden, C Murraydunning, L Thanabalasundaram, J Cowan, N Grace, G Cameron, Rosemary Drake, J B Phillips
    Abstract:

    Artificial tissues constructed from therapeutic Cells offer a promising approach for improving the treatment of severe peripheral nerve injuries. In this study the effectiveness of using CTX0E03, a conditionally immortalised human neural Stem Cell Line, as a source of allogeneic Cells for constructing living artificial nerve repair tissue was tested. CTX0E03 Cells were differentiated then combined with collagen to form engineered neural tissue (EngNT-CTX), stable aligned sheets of Cellular hydrogel. EngNT-CTX sheets were delivered within collagen tubes to repair a 12 mm sciatic nerve injury model in athymic nude rats. Autologous nerve grafts (autografts) and empty tubes were used for comparison. After 8 weeks functional repair was assessed using electrophysiology. Further, detailed histological and electron microscopic analysis of the repaired nerves was performed. Results indicated that EngNT-CTX supported growth of neurites and vasculature through the injury site and facilitated reinnervation of the target muscle. These findings indicate for the first time that a clinically validated allogeneic neural Stem Cell Line can be used to construct EngNT. This provides a potential 'off the shelf' tissue engineering solution for the treatment of nerve injury, overcoming the limitations associated with nerve autografts or the reliance on autologous Cells for populating repair constructs.

  • investigation of content stoichiometry and transfer of mirna from human neural Stem Cell Line derived exosomes
    PLOS ONE, 2016
    Co-Authors: Lara Stevanato, L Thanabalasundaram, Nickolai Vysokov, John Sinden
    Abstract:

    Exosomes are small (30-100 nm) membrane vesicles secreted by a variety of Cell types and only recently have emerged as a new avenue for Cell-to-Cell communication. They are natural shuttles of RNA and protein cargo, making them attractive as potential therapeutic delivery vehicles. MicroRNAs (miRNAs) are short non-coding RNAs which regulate biological processes and can be found in exosomes. Here we characterized the miRNA contents of exosomes derived from human neural Stem Cells (hNSCs). Our investigated hNSC Line is a clonal, conditionally immortalized Cell Line, compliant with good manufacturing practice (GMP), and in clinical trials for stroke and critical limb ischemia in the UK (clinicaltrials.gov: NCT01151124, NCT02117635, and NCT01916369). By using next generation sequencing (NGS) technology we identified the presence of a variety of miRNAs in both exosomal and Cellular preparations. Many of these miRNAs were enriched in exosomes indicating that Cells specifically sort them for extraCellular release. Although exosomes have been proven to contain miRNAs, the copy number quantification per exosome of a given miRNA remains unclear. Herein we quantified by real-time PCR a highly shuttled exosomal miRNA subtype (hsa-miR-1246) in order to assess its stoichiometry per exosome. Furthermore, we utilized an in vitro syStem to confirm its functional transfer by measuring the reduction in luciferase expression using a 3' untranslated region dual luciferase reporter assay. In summary, NGS analysis allowed the identification of a unique set of hNSC derived exosomal miRNAs. Stoichiometry and functional transfer analysis of one of the most abundant identified miRNA, hsa-miR-1246, were measured to support biological relevance of exosomal miRNA delivery.

  • a conditionally immortal clonal Stem Cell Line from human cortical neuroepithelium for the treatment of ischemic stroke
    Experimental Neurology, 2006
    Co-Authors: Kenneth Pollock, Paul Stroemer, Sara Patel, Lara Stevanato, Andrew Hope, Erik Miljan, Ziping Dong, Helen Hodges, Jack Price
    Abstract:

    Abstract Transplantation of neural Stem Cells into the brain is a novel approach to the treatment of chronic stroke disability. For clinical application, safety and efficacy of defined, stable Cell Lines produced under GMP conditions are required. To this end, a human neural Stem Cell Line, CTX0E03, was derived from human somatic Stem Cells following genetic modification with a conditional immortalizing gene, c -mycER TAM . This transgene generates a fusion protein that stimulates Cell proliferation in the presence of a synthetic drug 4-hydroxy-tamoxifen (4-OHT). The Cell Line is clonal, expands rapidly in culture (doubling time 50–60 h) and has a normal human karyotype (46 XY). In the absence of growth factors and 4-OHT, the Cells undergo growth arrest and differentiate into neurons and astrocytes. Transplantation of CTX0E03 in a rat model of stroke (MCAo) caused statistically significant improvements in both sensorimotor function and gross motor asymmetry at 6–12 weeks post-grafting. In addition, Cell migration and long-term survival in vivo were not associated with significant Cell proliferation. These data indicate that CTX0E03 has the appropriate biological and manufacturing characteristics necessary for development as a therapeutic Cell Line.

Rabindra N Padhy - One of the best experts on this subject based on the ideXlab platform.

  • human umbilical cord blood derived neural Stem Cell Line as a screening model for toxicity
    Neurotoxicity Research, 2017
    Co-Authors: Rajashree Patnaik, Rabindra N Padhy
    Abstract:

    The aim was to investigate whether a human neural Stem Cell (NSC) Line derived from human umbilical cord blood (hUCB) can be used for toxicity study. Toxicity of both neurotoxic environmental xenobiotics, methyl mercury chloride (CH3HgCl), lead acetate (CH3COOPb), and chlorpyrifos (CP), and non-neurotoxic insecticide, dichlorvos, as well as non-neurotoxic drugs, theophylLine and acetaminophen were assessed. Additionally, differentiation of neuronal and glial Cell Lines derived from hUCB was elucidated. It was observed that CH3HgCl was more toxic to human NSCs in comparison to CH3COOPb and CP. The minimum inhibitory concentration (MIC) value against NSCs was 3, 10, and 300 mg/L, in each staining process, acridine orange/ethidium bromide (AO/EB) staining, 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay, and Hoechst staining, for CH3HgCl, CP, and CH3COOPb, respectively. CH3HgCl had the LC25 value as 10.0, 14.4, and 12.7 mg/L, by staining method mentioned in succession. CP had the LC25 value as 21.9, 23.7, and 18.4 mg/L; similarly, CH3COOPb had LC25 values, successively as 616.9, 719.2, and 890.3 mg/L. LC50 values ranged from 18.2 to 21.7 mg/L for CH3HgCl, 56.4 to 60.2 mg/L for CP, and 1000 to 1460.1 for CH3COOPb. TheophylLine, acetaminophen, and dichlorvos had no impact on the viability of NSCs. This work justified that hUCB-NSC model can be used for toxicity study.