Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep

Abstract

Mesenchymal stem cells are multipotent cells that can be isolated from adult bone marrow and can be induced in vitro and in vivo to differentiate into a variety of mesenchymal tissues, including bone, cartilage, tendon, fat, bone marrow stroma, and muscle1,2. Despite their potential clinical utility for cellular and gene therapy, the fate of mesenchymal stem cells after systemic administration is mostly unknown. To address this, we transplanted a well-characterized human mesenchymal stem cell population3 into fetal sheep early in gestation, before and after the expected development of immunologic competence. In this xenogeneic system, human mesenchymal stem cells engrafted and persisted in multiple tissues for as long as 13 months after transplantation. Transplanted human cells underwent site-specific differentiation into chondrocytes, adipocytes, myocytes and cardiomyocytes, bone marrow stromal cells and thymic stroma. Unexpectedly, there was long-term engraftment even when cells were transplanted after the expected development of immunocompetence. Thus, mesenchymal stem cells maintain their multipotential capacity after transplantation, and seem to have unique immunologic characteristics that allow persistence in a xenogeneic environment. Our data support the possibility of the transplantability of mesenchymal stem cells and their potential utility in tissue engineering, and cellular and gene therapy applications.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Screening of sheep tissues (n = 29) after transplantation, by DNA PCR using probes specific for human β-2 microglobulin sequences.
Figure 2: Immunohistochemistry with human-specific antibody against β-2 microglobulin.
Figure 3: Evidence for human cell differentiation.
Figure 4: Evidence for cardiac and skeletal myocyte differentiation.

Similar content being viewed by others

References

  1. Caplan, A.I. The mesengenic process. Clin. Plastic Surg. 21, 429–435 (1994).

    CAS  Google Scholar 

  2. Prockop, D.J. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276, 71–74 (1997).

    Article  CAS  Google Scholar 

  3. Pittenger, M.F., et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999).

    Article  CAS  Google Scholar 

  4. Zanjani, E.D., Flake, A.W., Rice, H., Hedrick, M. & Tavassoli, M. Long-term repopulating ability of xenogeneic transplanted human fetal liver hematopoietic stem cells in sheep. J. Clin. Invest. 93, 1051–1055 (1994).

    Article  CAS  Google Scholar 

  5. Silverstein, A.M., Prendergast, R.A. & Kraner, K.L. Fetal response to antigenic stimulus IV. Rejection of skin homografts by the fetal lamb. J. Exp. Med. 119, 955–964 (1964).

    Article  CAS  Google Scholar 

  6. Flake, A.W., Harrison, M.R., Adzick, N.S. & Zanjani, E.D. Transplantation of fetal hematopoietic stem cells in utero: the creation of hematopoietic chimeras. Science 233, 776–778 (1986).

    Article  CAS  Google Scholar 

  7. Zanjani, E.D., Ascensao, J.L. & Tavassoli, M. Liver-derived fetal hematopoietic stem cells selectively and preferentially home to the fetal bone marrow. Blood 81, 399–404 (1993).

    CAS  PubMed  Google Scholar 

  8. Zanjani, E., Almeida-Porada, G., Ascensao, J., MacKintosh, F. & Flake, A. Transplantation of hematopoietic stem cells in utero. Stem Cells 15, 79–93 (1997).

    Article  Google Scholar 

  9. Zanjani, E.D., Almeida-Porada, G. & Flake, A.W. The human/sheep xenograft model: a large animal model of human hematopoiesis. Int. J. Hematol. 63, 179–192 (1996).

    Article  CAS  Google Scholar 

  10. Zanjani, E.D., Almeida-Porada, G., Livingston, A.G., Flake, A.W. & Ogawa, M. Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. Exp. Hematol. 26, 353–360 (1998).

    CAS  PubMed  Google Scholar 

  11. Fourcade, C. et al. Expression of CD23 by human bone marrow stromal cells. Eur. Cytokine Network 3, 539–543 (1992).

    CAS  Google Scholar 

  12. Schlossman, S., Bloumsell, L. & Gilks, W. in Leukocyte Typing V: White Cell Differentiation Antigens. (Oxford University Press, New York, 1995).

    Google Scholar 

  13. Sharp, A.H. et al. Differential immunohistochemical localization of inositol 1,4,5-trisphosphate- and ryanodine-sensitive Ca2+ release channels in rat brain. J. Neurosci. 13, 3051–3063 (1993).

    Article  CAS  Google Scholar 

  14. Nicholson, L.V. et al. Dystrophin or a “related protein” in Duchenne muscular dystrophy? Acta. Neurol. Scand. 86, 8–14 (1992).

    Article  CAS  Google Scholar 

  15. Keating, A. et al. Donor origin of the in vitro haematopoietic microenvironment after marrow transplantation in man. Nature 298, 280–283 (1982).

    Article  CAS  Google Scholar 

  16. Anklesaria, P. et al. Engraftment of a clonal bone marrow stromal cell line in vivo stimulates hematopoietic recovery from total body irradiation. Proc. Natl. Acad. Sci. USA 84, 7681–7685 (1987).

    Article  CAS  Google Scholar 

  17. Ferrari, G. et al. Muscle regeneration by bone marrow-derived myogenic progenitors. Science 279, 1528–1530 (1998).

    Article  CAS  Google Scholar 

  18. Horwitz, E.M. et al. Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfecta. Nature Med. 5, 309–313 (1999).

    Article  CAS  Google Scholar 

  19. Hou, Z. et al. Osteoblast-specific gene expression after transplantation of marrow cells: implications for skeletal gene therapy. Proc. Natl. Acad. Sci. USA 96, 7294–7299 (1999).

    Article  CAS  Google Scholar 

  20. Gussoni, E. et al. Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature 401, 390–394 (1999).

    CAS  Google Scholar 

  21. Pereira, R.F. et al. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc. Natl. Acad. Sci. USA 92, 4857–4861 (1995).

    Article  CAS  Google Scholar 

  22. Pereira, R.F. et al. Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta. Proc. Natl. Acad. Sci. USA 95, 1142–1147 (1998).

    Article  CAS  Google Scholar 

  23. Culling, C.F.A. in Handbook of Histopathological and Histochemical Techniques (Butterworth and Co., London, 1974).

    Google Scholar 

  24. Gilliland, G., Perrin, S., Blanchard, K. & Bunn, F. Analysis of cytokine mRNA and DNA: detection and quantification by competetive polymerase chain reaction. Proc. Natl. Acad. Sci. USA 87, 2725–2729 (1990).

    Article  CAS  Google Scholar 

  25. Van Der Loos, C.M., Becker, A.E. & Van Den Oord, J.J. Practical suggestions for successful immunoenzyme double-staining experiments. Histochem. J. 25, 1–11 (1993).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Alan W. Flake.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liechty, K., MacKenzie, T., Shaaban, A. et al. Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. Nat Med 6, 1282–1286 (2000). https://doi.org/10.1038/81395

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/81395

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing