1st Page



This article in JAS

  1. Vol. 89 No. 11, p. 3460-3472
    Received: Jan 20, 2011
    Accepted: June 14, 2011
    Published: December 4, 2014

    3 Corresponding author(s):


Toward an ideal animal model to trace donor cell fates after stem cell therapy: Production of stably labeled multipotent mesenchymal stem cells from bone marrow of transgenic pigs harboring enhanced green fluorescence protein gene1

  1. F. S. H. Hsiao*†22,
  2. W. S. Lian*22,
  3. S. P. Lin†‡§#22,
  4. C. J. Lin*,
  5. Y. S. Lin*,
  6. E. C. H. Cheng*,
  7. C. W. Liu*,
  8. C. C. Cheng,
  9. P. H. Cheng*,
  10. S. T. Ding*†‖,
  11. K. H. Lee,
  12. T. F. Kuo**,
  13. C. F. Cheng††‡‡,
  14. W. T. K. Cheng 3 and
  15. S. C. Wu 3
  1. Department of Animal Science and Technology, and
    Institute of Biotechnology, National Taiwan University, Taipei 106 Taiwan;
    Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115 Taiwan;
    Research Center for Developmental Biology and Regenerative Medicine,
    Center for Systems Biology, and
    Center for Biotechnology, Department of Animal Science and Technology, National Taiwan University, Taipei 106 Taiwan;
    Division of Biotechnology, Animal Technology Institute Taiwan, Miaoli 35053 Taiwan;
    Department of Veterinary Medicine, National Taiwan University, Taipei 106 Taiwan;
    Institute of Biomedical Science, Academia Sinica, Taipei 115 Taiwan;
    Department of Medical Research, Tzu Chi General Hospital and Department of Pediatrics, Tzu Chi University, Hualien 97004 Taiwan; and
    Department of Animal Science and Biotechnology, Tunghai University, Taichung 407 Taiwan



The discovery of postnatal mesenchymal stem cells (MSC) with their general multipotentiality has fueled much interest in the development of cell-based therapies. Proper identification of transplanted MSC is crucial for evaluating donor cell distribution, differentiation, and migration. Lack of an efficient marker of transplanted MSC has precluded our understanding of MSC-related regenerative studies, especially in large animal models such as pigs. In the present study, we produced transgenic pigs harboring an enhanced green fluorescent protein (EGFP) gene. The pigs provide a reliable and reproducible source for obtaining stable EGFP-labeled MSC, which is very useful for donor cell tracking after transplantation. The undifferentiated EGFP-tagged MSC expressed a greater quantity of EGFP while maintaining MSC multipotentiality. These cells exhibited homogeneous surface epitopes and possessed classic trilineage differentiation potential into osteogenic, adipogenic, and chondrogenic lineages, with robust EGFP expression maintained in all differentiated progeny. Injection of donor MSC can dramatically increase the thickness of infarcted myocardium and improve cardiac function in mice. Moreover, the MSC, with their strong EGFP expression, can be easily distinguished from the background autofluorescence in myocardial infarcts. We demonstrated an efficient, effective, and easy way to identify MSC after long-term culture and transplantation. With the transgenic model, we were able to obtain stem or progenitor cells in earlier passages compared with the transfection of traceable markers into established MSC. Because the integration site of the transgene was the same for all cells, we lessened the potential for positional effects and the heterogeneity of the stem cells. The EGFP-transgenic pigs may serve as useful biomedical and agricultural models of somatic stem cell biology.

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