Use of Bioluminescent Imaging to Assay the Transplantation of Immortalized Human Fetal Hepatocytes into Mice

Cell Transplantation

Article

Alexander D. Borowsky¹  Sanjeev Gupta²  Sanjiv S. Gambhir³  Andreea M. Catana⁴  Mark A. Zern⁴  Jian Wu⁴  Young Seok Kim⁴  Sang Jeong Yoon⁴  Moon Seok Choi⁵ 

[1] Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710, Korea;Marion Bessin Liver Research Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY, USA;Molecular Imaging Program at Stanford, Departments of Radiology and Bioengineering, Bio-X Program, Stanford University, Stanford, CA, USA;Transplant Research Institute, UC Davis Medical Center, Sacramento, CA, USA;Transplant Research Institute, UC Davis Medical Center, Sacramento, CA, USA;Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710, Korea;

关键词: Hepatocytes;    Stem cells;    Transplantation;    Luciferase;    Imaging techniques;   

DOI  :  10.3727/096368908786576471

 received in 2007-06-11, accepted in 2008-01-25,  发布年份 2008

来源: Sage Journals

【 摘 要 】

Noninvasive serial monitoring of the fate of transplanted cells would be invaluable to evaluate the potential therapeutic use of human hepatocyte transplantation. Therefore, we assessed the feasibility of bioluminescent imaging using double or triple fusion lentiviral vectors in a NOD-SCID mouse model transplanted with immortalized human fetal hepatocytes. Lentiviral vectors driven by the CMV promoter were constructed carrying reporter genes: firefly luciferase and green fluorescence protein with or without herpes simplex virus type 1 thymidine kinase. Human fetal hepatocytes immortalized by telomerase reconstitution (FH-hTERT) were successfully transduced with either of these fusion vectors. Two million stably transduced cells selected by fluorescence-activated cell sorting were injected into the spleens of NOD-SCID mice pre-treated with methylcholanthrene and monocrotaline. The transplanted mice were serially imaged with a bioluminescence charged-coupled device camera after D-luciferin injection. Bioluminescence signal intensity was highest on day 3 (6.10 ± 2.02 × 105 p/s/cm2/sr, mean ± SEM), but decreased to 2.26 ± 1.54 × 105 and 7.47 ± 3.09 × 104 p/s/cm2/sr on day 7 and 10, respectively (p = 0.001). ELISA for human albumin in mice sera showed that levels were similar to those of control mice on day 2 (3.25 ± 0.92 vs. 2.84 ± 0.59 ng/ml, mean ± SEM), peaked at 18.04 ±3.11 ng/ml on day 7, and decreased to 8.93 ± 1.40 and 3.54 ± 0.87 ng/ml on day 14 and 21, respectively (p = 0.02). Real-time quantitative RT-PCR showed gene expression levels of human albumin, α1-antitrypsin, and transferrin in mouse liver were 60.7 ± 6.5%, 26.0 ± 1.4%, and 156.8 ± 62.4% of those of primary human adult hepatocytes, respectively, and immunohistochemistry revealed cells with human albumin and α1-antitrypsin expression in the mouse liver. In conclusion, our study demonstrated that bioluminescent imaging appears to be a sensitive, noninvasive modality for serial monitoring of transplanted hepatic stem cells.

【 授权许可】

Unknown   
© 2008 Cognizant Comm. Corp.