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dc.contributor.authorCreane, Michael
dc.contributor.authorHoward, Linda
dc.contributor.authorO'Brien, Timothy
dc.contributor.authorColeman, Cynthia M.
dc.identifier.citationCreane, Michael, Howard, Linda, O'Brien, Timothy, & Coleman, Cynthia M. (2017). Biodistribution and retention of locally administered human mesenchymal stromal cells: Quantitative polymerase chain reaction–based detection of human DNA in murine organs. Cytotherapy, 19(3), 384-394. doi:
dc.description.abstractBackground. Determining the distributive fate and retention of a cell therapy product after administration is an essential part of characterizing it's biosafety profile. Therefore, regulatory guidelines stipulate that biodistribution assays are a requirement prior to advancing a cell therapy to the clinic. Here the development of a highly sensitive quantitative polymerase chain reaction (qPCR)-based method of tracking the biodistribution and retention of human mesenchymal stromal cells (hMSCs) in mice, rats or rabbits is described. Methods. A primer-probe based qPCR assay was developed to detect and quantify human Alu sequences in a heterogeneous sample of human DNA (hDNA) and murine DNA from whole organ genomic DNA extracts. The assay measures the amount of genomic hDNA by amplifying a 31 base pair sequence of the human Alu (hAlu) repeat sequence, thus enabling the detection of 0.1 human cells in 1.5 x 10(6) heterogeneous cells. Results. Using this assay we investigated the biodistribution of 3 x 10(5) intramuscularly injected hMSCs in Balb/c nude mice. Genomic DNA was extracted from murine organs and hAlu sequences were quantified using qPCR analysis. After 3 months, hDNA ranging from 0.07%-0.58% was detected only at the injection sites and not in the distal tissues of the mice. Discussion. This assay represents a reproducible, sensitive a method of detecting hDNA in rodent and lapine models.This manuscript describes the method employed to generate preclinical biodistribution data that was accepted by regulatory bodies in support of a clinical trial application.en_IE
dc.description.sponsorshipThis work was supported by REDDSTAR: European Union Seventh Framework Programme HEALTH-F2-2012-305736, Science Foundation Ireland Strategic Research Cluster Grant No. SFI:09/SRC B1794 and the European Regional Development Fund. The authors acknowledge the facilities as well as the scientific and technical assistance of the National Centre for Biomedical Engineering Science Genomics Facility at the National University of Ireland Galway, a facility that is funded by National University of Ireland Galway and the Irish Government's Programme for Research in Third Level Institutions, Cycles 4 and 5, National Development Plan 2007–2013. hMSCs used in this study were manufactured according to GMP by the Centre For Cell Manufacturing Ireland at the National University of Ireland in Galway.en_IE
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Ireland
dc.subjectcell therapyen_IE
dc.subjectgenomic DNAen_IE
dc.subjecthuman Alu sequenceen_IE
dc.subjectmesenchymal stromal cellen_IE
dc.subjectpolymerase chain reactionen_IE
dc.subjecttranslational stem cell researchen_IE
dc.subjectREAL-TIME PCRen_IE
dc.titleBiodistribution and retention of locally administered human mesenchymal stromal cells: Quantitative polymerase chain reaction–based detection of human DNA in murine organsen_IE
dc.contributor.funderSeventh Framework Programmeen_IE
dc.contributor.funderScience Foundation Irelanden_IE
dc.local.contactLinda Howard, Biomedical Science Building, Dan, Nui Galway. 5268 Email:
dcterms.projectinfo:eu-repo/grantAgreement/EC/FP7::SP1::HEALTH/305736/EU/Repair of Diabetic Damage by Stromal Cell Administration/REDDSTARen_IE
dcterms.projectinfo:eu-repo/grantAgreement/SFI/SFI Strategic Research Cluster/09/SRC/B1794/IE/SRC REMEDI: REMEDI-2: From Research to Human Therapy/en_IE

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