Polycistronic lentivirus induced pluripotent stem cells from skin biopsies after long term storage,blood outgrowth endothelial cells and cells from milk teeth |
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Authors: | C Dambrot S van de Pas L van Zijl B Brändl JW Wang MJ Schalij RC Hoeben DE Atsma HM Mikkers CL Mummery C Freund |
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Institution: | 1. Department of Anatomy, Leiden University Medical Centre, Leiden, The Netherlands;2. Department of Cardiology, Leiden University Medical Centre, Leiden, The Netherlands;3. Department of Molecular Cell Biology, Leiden University Medical Centre, Leiden, The Netherlands;4. Zentrum für Integrative Psychiatrie, University Hospital Schleswig–Holstein, Kiel, Germany;5. Einthoven Laboratory for Experimental Vascular Medicine, Department of Thrombosis and Hemostasis, Leiden University Medical Centre, Leiden, The Netherlands;6. Laboratory of Experimental Cardiology, University Medical Centre Utrecht, Utrecht, The Netherlands |
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Abstract: | The generation of human induced pluripotent stem cells (hiPSCs) requires the collection of donor tissue, but clinical circumstances in which the interests of patients have highest priority may compromise the quality and availability of cells that are eventually used for reprogramming. Here we compared (i) skin biopsies stored in standard physiological salt solution for up to two weeks (ii) blood outgrowth endothelial cells (BOECs) isolated from fresh peripheral blood and (iii) children's milk teeth lost during normal replacement for their ability to form somatic cell cultures suitable for reprogramming to hiPSCs. We derived all hiPSC lines using the same reprogramming method (a conditional (FLPe) polycistronic lentivirus) and under similar conditions (same batch of virus, fetal calf serum and feeder cells). Skin fibroblasts could be reprogrammed robustly even after long-term biopsy storage. Generation of hiPSCs from juvenile dental pulp cells gave similar high efficiencies, but that of BOECs was lower. In terms of invasiveness of biopsy sampling, biopsy storage and reprogramming efficiencies skin fibroblasts appeared best for the generation of hiPSCs, but where non-invasive procedures are required (e.g. for children and minors) dental pulp cells from milk teeth represent a valuable alternative. |
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Keywords: | hESC human embryonic stem cells (h)iPSC (human) induced pluripotent stem cell BOECs blood outgrowth endothelial cells DMEM Dulbecco's Modified Eagle Media PBS phosphate buffered saline MEF mouse embryonic fibroblast AP alkaline phosphatase MOI multiplicity of infection NEAA non-essential amino acids FCS fetal calf serum KOSR knock-out serum replacement AFP α-fetoprotein |
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