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dc.contributor.advisorLam, Wilbur A.
dc.contributor.authorTran, Reginald
dc.date.accessioned2018-01-22T21:05:26Z
dc.date.available2018-01-22T21:05:26Z
dc.date.created2016-12
dc.date.issued2016-11-15
dc.date.submittedDecember 2016
dc.identifier.urihttp://hdl.handle.net/1853/59163
dc.description.abstractEx vivo gene therapy using lentiviral vectors (LVs) is a proven approach to treat and potentially cure many hematologic disorders and cancer, but remains stymied by cumbersome, cost-prohibitive, and scale-limited production processes that cannot meet the demands of current clinical protocols for widespread implementation. In this work, we describe the development, characterization, and application of a microfluidic, mass transport-based approach that overcomes the diffusion limitations of current transduction platforms to enhance LV gene transfer kinetics and efficiency. This novel ex vivo LV transduction platform is flexible in design, easy to use, scalable, and compatible with standard cell culture transduction reagents and LV preparations as it mechanistically relies solely on physical principles. Hematopoietic cell lines, primary human T cells, and primary hematopoietic stem and progenitor cells were used to assess microfluidic transduction, which was demonstrated to occur up to 5-fold faster and required as little as ~1/20th of lentivirus needed in conventional clinical transduction protocols. In vivo application of microfluidics using hematopoietic stem and progenitor cells of C57BL/6J hemophilia A mice transduced with factor VIII-encoding LV and transplanted into hemophilic donors demonstrated that LV usage and transduction time can significantly be reduced with microfluidics, which can be incorporated with other transduction enhancement strategies. Moreover, when using equal amounts of vector, only hemophiliac mice transplanted with microfluidic-transduced Sca-1+ cells were able to restore plasma fVIII levels to normal, demonstrating the greater efficiency of the microfluidic transduction platform. Overall, this work highlights the utility of microfluidics to overcome diffusion limitations of standard cell culture systems to improve efficiency and utilization of LVs for clinical gene therapy.
dc.format.mimetypeapplication/pdf
dc.language.isoen_US
dc.publisherGeorgia Institute of Technology
dc.subjectMicrofluidics
dc.subjectGene therapy
dc.subjectHemophilia
dc.subjectHematology
dc.subjectCell manufacturing
dc.subjectCancer immunotherapy
dc.subjectHematopoietic stem cells
dc.subjectT cells
dc.subjectMass transport
dc.subjectLentivirus
dc.titleA microfluidics-based paradigm for clinical lentivector gene transfer
dc.typeDissertation
dc.description.degreePh.D.
dc.contributor.departmentBiomedical Engineering (Joint GT/Emory Department)
thesis.degree.levelDoctoral
dc.contributor.committeeMemberDixon, J. Brandon
dc.contributor.committeeMemberDoering, Christopher B.
dc.contributor.committeeMemberLe Doux, Joseph M.
dc.contributor.committeeMemberSulchek, Todd
dc.date.updated2018-01-22T21:05:26Z


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