Expressing humanized bacterial luciferase in stem cells: Moving beyond fireflyluciferase to expand the informational capacity of animal models for regenerativemedicine

Expressing humanized bacterial luciferase in stem cellsMoving beyond firefly luciferase toexpand the informational capacity of animal models for regenerative medicine Project Summary This Small Business Technology TransferSTTRPhase II project proposes to develop complementary autonomously bioluminescentautobioluminescentin vitro stem cell lines and in vivo small animal model systems that enable the continuousreagent freeand real time bioimaging of mesenchymal stem cellMSClocalizationdifferentiation into adipocytechondrocyteand osteocyte lineagesand persistence postdifferentiation at the site of activationThese models will specifically address the National Institutes of Health s request for new techniques for non invasivelong term tracking of stem cell survivabilityengraftmentand migration following in vivo implantationBy addressing this critical need for new methods capable of elucidating the mechanisms underlying how stem cells identify areas of dysfunction within the bodydifferentiate into the relevant tissues required to correct the maladyand persist in synergy with existing tissue to enable long term functionalitythese tools will significantly improve the transition of regenerative medicine studies towards translational and clinical practice outcomesThe autobioluminescent MSCs developed byBioTech under our Phase I effort demonstrated the ability to track MSC localization in vitro and in vivo similarly to existing optical imaging approachesbut with significantly reduced cost and personnel effortFurthermorethese models also negated the need for sample destruction or the stressful and potentially influential injection of an activating chemical concurrent with imaging while simultaneously providing an uninterrupted stream of visual data over the lifetime of the reporter cell as it interacts with its environment and undergoes differentiationIn partnership with the University of Tennessee Medical Centerthis proposal will expand upon these accomplishments to develop fully self contained autobioluminescent MSC based cellular models capable of specifically reporting on their differentiation into adipocytechondrocyteand osteocyte lineagesand complementary small animal models harboring native MSCs genetically programmed to autonomously enact their reporter functionality only following differentiation into adipocytechondrocyteor osteocyte lineages in response to wounding or exogenous stimulationThese models will overcome the primary technical hurdles encountered with all existing bioluminescent and fluorescent stem cells currently on the market from companies such as PerkinElmerThermoFisher Life TechnologiesPromegaandsmaller specialized business entities in the U Salonewhich comprise an estimated market value of at least $ Bwith a predicated annual growth rate ofWe believe that the products developed in this effort will be capable of significantly improving the throughput and effectiveness of regenerative medicine studies and advancing our understanding of stem cell based treatment efficiency and efficacy to improve both public health and consumer safetyThe functional demonstrations and data gathered in this effort will position these models to thrive within this market and produce an immediate and significant impact on the field of regenerative medicine that will benefit the population at large