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Topics {✒️}

sai kiang lim multipotent stem cells surface antigen profile de kleijn dp open access article genome-wide gene expression mesenchymal stem cells human esc-derived hues9 human esc-derived mscs stem cell research ruenn chai lai retrovirus-mediated gene transfer gene expression profile chuen neng lee fatih arslan article download pdf cell-based treatment modality myocardial ischemia/reperfusion injury graft-versus-host-disease [1] pluripotent stem cells intestinal stem cells full size image stem cell crypt saline-treated control group fetal mscs immortalized stromal-derived factor-1 hplc protein profile stem cell res cell-based therapy infinite supply onco-protein unlike oncogene autologous cell therapy author information authors gfp-transfected cells progressed unlike cell-based therapies secreted bi-lipid vesicles cardiac regenerative therapy myc-transformed cells continued mrna/protein expression analysis mesenchymal cell types uv-visible detector operated related subjects myc-transformed cells expressed myocardial ischemia/reperfusion esc-derived mscs chemically defined medium relative myc-transcript level privacy choices/manage cookies stem cells myocardial infarct size

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         headline:Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs
         description:Exosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs) have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes. The hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining. MYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved. Our results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time for cell production and thereby reduces production costs.
         datePublished:2011-04-25T00:00:00Z
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            Infarct Size
            Infinite Supply
            Surface Antigen Profile
            Exosome Treatment
            Biomedicine
            general
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      headline:Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs
      description:Exosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs) have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes. The hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining. MYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved. Our results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time for cell production and thereby reduces production costs.
      datePublished:2011-04-25T00:00:00Z
      dateModified:2011-04-25T00:00:00Z
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      pageEnd:10
      license:https://creativecommons.org/licenses/by/2.0
      sameAs:https://doi.org/10.1186/1479-5876-9-47
      keywords:
         Conditioned Medium
         Infarct Size
         Infinite Supply
         Surface Antigen Profile
         Exosome Treatment
         Biomedicine
         general
         Medicine/Public Health
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      name:Institute of Medical Biology, A*STAR, Singapore
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      name:Interuniversity Cardiology Institute of the Netherlands, Utrecht, the Netherlands
      name:Institute of Medical Biology, A*STAR, Singapore
      name:Department of Surgery, YLL School of Medicine, Singapore

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