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Transgenic mouse with a stably integrated reporter genes can be a valuable resource for obtaining uniformly labeled stem cells, tissues, and organs for various applications. We have generated a transgenic mouse model that ubiquitously expresses a tri-fusion reporter gene fluc2-tdTomato-ttk driven by a constitutive chicken β-actin promoter. This -Tri-Modality Reporter Mouse- system allows one to isolate most cells from this donor mouse and image them for bioluminescent fluc2, fluorescent tdTomato, and positron emission tomography PET ttk modalities. Transgenic colonies with different levels of tri-fusion reporter gene expression showed a linear correlation between all three-reporter proteins R2=0.89 for TdTomato vs Fluc, R2=0.94 for Fluc vs TTK, R2=0.89 for TdTomato vs TTK in vitro from tissue lysates and in vivo by optical and PET imaging. Mesenchymal stem cells MSCs isolated from this transgenics showed high level of reporter gene expression, which linearly correlated with the cell numbers R2=0.99 for bioluminescence imaging BLI. Both BLI R2=0.93 and micro-PET R2=0.94 imaging of the subcutaneous implants of Tri-Modality Reporter Mouse derived MSCs in nude mice showed linear correlation with the cell numbers and across different imaging modalities R2=0.97. Serial imaging of MSCs transplanted to mice with acute myocardial infarction MI by intramyocardial injection exhibited significantly higher signals in MI heart at days 2, 3, 4, and 7 p<0.01. MSCs transplanted to the ischemic hindlimb of nude mice showed significantly higher BLI and PET signals in the first 2 weeks that dropped by 4th week due to poor cell survival. However, laser Doppler perfusion imaging revealed that blood circulation in the ischemic limb was significantly improved in the MSCs transplantation group compared with the control group. In summary, this mouse can be used as a source of donor cells and organs in various research areas such as stem cell research, tissue engineering research, and organ transplantation.



Autor: Xinrui Yan, Pritha Ray, Ramasamy Paulmurugan, Ricky Tong, Yongquan Gong, Ataya Sathirachinda, Joseph C. Wu, Sanjiv S. Gambhir

Fuente: http://plos.srce.hr/



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