Advertisement

Stem Cell Reviews and Reports

, Volume 10, Issue 1, pp 79–85 | Cite as

The Potential Role of Genetically-Modified Pig Mesenchymal Stromal Cells in Xenotransplantation

  • Jiang Li
  • Mohamed B. Ezzelarab
  • David Ayares
  • David K. C. CooperEmail author
Article

Abstract

Mesenchymal stromal cells (MSCs) are known to have regenerative, anti-inflammatory, and immunodulatory effects. There are extensive indications that pig MSCs function satisfactorily across species barriers. Pig MSCs might have considerable therapeutic potential, particularly in xenotransplantation, where they have several potential advantages. (i) pMSCs can be obtained from the specific organ- or cell-source donor pig or from an identical (cloned) pig. (ii) They are easy to obtain in large numbers, negating the need for prolonged ex vivo expansion. (iii) They can be obtained from genetically-engineered pigs, and the genetic modification can be related to the therapeutic goal of the MSCs. We have reviewed our own studies on MSCs from genetically-engineered pigs, and summarize them here. We have successfully harvested and cultured MSCs from wild-type and genetically-engineered pig bone marrow and adipose tissue. We have identified several pig (p)MSC surface markers (positive for CD29, CD44, CD73, CD105, CD166, and negative for CD31, CD45), have demonstrated their proliferation and differentiation (into adipocytes, osteoblasts, and chondroblasts), and evaluated their antigenicity and immune suppressive effects on human peripheral blood mononuclear cells and CD4+T cells. They have identical or very similar characteristics to MSCs from other mammals. Genetically-modified pMSCs are significantly less immunogenic than wild-type pMSCs, and downregulate the human T cell response to pig antigens as efficiently as do human MSCs. We hypothesized that pMSCs can immunomodulate human T cells through induction of apoptosis or anergy, or cause T cell phenotype switching with induction of regulatory T cells, but we could find no evidence for these mechanisms. However, pMSCs upregulated the expression of CD69 on human CD4+ and CD8+ T cells, the relevance of which is currently under investigation. We conclude that MSCs from genetically-engineered pigs should continue to be investigated for their immunomodulatory (and regenerative and anti-inflammatory) effects in pig-to-nonhuman primate organ and cell transplantation models.

Keywords

Mesenchymal stromal cells Pig Xenotransplantation 

Abbreviations

AdMSC

Adipose-derived mesenchymal stromal cells

AECs

Aortic endothelial cells

Gal

Galactose-α1,3-galactose

GTKO

α1,3-galactosyltransferase gene-knockout

MLR

Mixed lymphocyte reaction

MSC

Mesenchymal stromal cells

PBMC

Peripheral blood mononuclear cells

SLA

Swine leukocyte antigen

Notes

Acknowledgments

Jiang Li is funded by the China Scholarship Council. Mohamed Ezzelarab is supported in part by the Joseph A. Patrick Research Fellowship in Transplantation of the Thomas E. Starzl Transplantation Institute of the University of Pittsburgh. Studies on xenotransplantation at the Thomas E. Starzl Transplantation Institute are supported in part by CMRF (competitive medical research funding) from the University of Pittsburgh Medical Center, NIH grants #1U19AI090959-01 and #1PO1 HL107152, and by Sponsored Research Agreements between the University of Pittsburgh and Revivicor, Inc., Blacksburg, VA.

Conflict of Interest

DA is an employee of Revivicor Inc., Blacksburg, VA. The other authors report no conflicts of interest.

References

  1. 1.
    Friedenstein, A. J., Petrakova, K. V., Kurolesova, A. I., & Frolova, G. P. (1968). Heterotopic of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation, 6, 230–247.PubMedCrossRefGoogle Scholar
  2. 2.
    Sordi, V. (2009). Mesenchymal stem cell homing capacity. Transplantation, 87, S42–S45.PubMedCrossRefGoogle Scholar
  3. 3.
    Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., et al. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8, 315–317.PubMedCrossRefGoogle Scholar
  4. 4.
    Wang, L., Zhao, Y., & Shi, S. (2012). Interplay between mesenchymal stem cells and lymphocytes: implications for immunotherapy and tissue regeneration. Journal of Dental Research, 91, 1003–1010.PubMedCrossRefGoogle Scholar
  5. 5.
    Wang, S., Qu, X., & Zhao, R. C. (2012). Clinical applications of mesenchymal stem cells. Journal of Hematology and Oncology, 5, 19.PubMedCrossRefGoogle Scholar
  6. 6.
    Le Blanc, K., Frassoni, F., Ball, L., Locatelli, F., Roelofs, H., Lewis, I., et al. (2008). Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet, 371, 1579–1586.PubMedCrossRefGoogle Scholar
  7. 7.
    Le Blanc, K., Rasmusson, I., Gotherstrom, C., Seidel, C., Sundberg, B., Sundin, M., et al. (2004). Mesenchymal stem cells inhibit the expression of CD25 (interleukin-2 receptor) and CD38 on phytohaemagglutinin-activated lymphocytes. Scandinavian Journal of Immunology, 60, 307–315.PubMedCrossRefGoogle Scholar
  8. 8.
    Reinders, M. E., Fijter, J. W., Roelofs, H., Bajema, I. M., Vries, D. K., Schaapherder, A. F., et al. (2013). Autologous bone marrow-derived mesenchymal stromal cells for the treatment of allograft rejection after renal transplantation: results of a phase I study. Stem Cells Translational Medicine, 2, 107–111.PubMedCrossRefGoogle Scholar
  9. 9.
    Mesenchymal stem cell transplantation in the treatment of chronic allograft nephropathy. (Under investigation, ClinicalTrials.gov Identifier: NCT00659620)Google Scholar
  10. 10.
    Perico, N., Casiraghi, F., Introna, M., Gotti, E., Todeschini, M., Cavinato, R. A., et al. (2011). Autologous mesenchymal stromal cells and kidney transplantation: a pilot study of safety and clinical feasibility. Clinical Journal of the American Society of Nephrology, 6, 412–422.PubMedCrossRefGoogle Scholar
  11. 11.
    Li, J., Ezzelarab, M. B., & Cooper, D. K. (2012). Do mesenchymal stem cells function across species barriers? Relevance for xenotransplantation. Xenotransplantation, 19, 273–285.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Ezzelarab, M., Ayares, D., & Cooper, D. K. (2010). The potential of genetically-modified pig mesenchymal stromal cells in xenotransplantation. Xenotransplantation, 17, 3–5.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Ezzelarab, M., Ezzelarab, C., Wilhite, T., Kumar, G., Hara, H., Ayares, D., et al. (2011). Genetically-modified pig mesenchymal stromal cells: xenoantigenicity and effect on human T-cell xenoresponses. Xenotransplantation, 18, 183–195.PubMedCrossRefGoogle Scholar
  14. 14.
    Kumar, G., Hara, H., Long, C., Shaikh, H., Ayares, D., Cooper, D. K., et al. (2012). Adipose-derived mesenchymal stromal cells from genetically modified pigs: immunogenicity and immune modulatory properties. Cytotherapy, 14, 494–504.PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Li, J., Andreyeva, O., Chen, M., Marcoa, M., Iwasea, H., Long, C., et al. (2013). Human T cells upregulate CD69 after coculture with xenogeneic genetically-modified pig mesenchymal stromal cells. Cellular Immunology (in press).Google Scholar
  16. 16.
    Cooper, D. K., Ekser, B., Burlak, C., Ezzelarab, M., Hara, H., Paris, L., et al. (2012). Clinical lung xenotransplantation–what donor genetic modifications may be necessary? Xenotransplantation, 19, 144–158.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Ekser, B., Ezzelarab, M., Hara, H., van der Windt, D. J., Wijkstrom, M., Bottino, R., et al. (2012). Clinical xenotransplantation: the next medical revolution? Lancet, 379, 672–683.PubMedCrossRefGoogle Scholar
  18. 18.
    Hara, H., Long, C., Lin, Y. J., Tai, H. C., Ezzelarab, M., Ayares, D., et al. (2008). In vitro investigation of pig cells for resistance to human antibody-mediated rejection. Transplant International, 21, 1163–1174.PubMedCrossRefGoogle Scholar
  19. 19.
    Phelps, C. J., Koike, C., Vaught, T. D., Boone, J., Wells, K. D., Chen, S. H., et al. (2003). Production of alpha 1,3-galactosyltransferase-deficient pigs. Science, 299, 411–414.PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Brusko, T. M. (2009). Mesenchymal stem cells: a potential border patrol for transplanted islets? Diabetes, 58, 1728–1729.PubMedCrossRefGoogle Scholar
  21. 21.
    Solari, M. G., Srinivasan, S., Boumaza, I., Unadkat, J., Harb, G., Garcia-Ocana, A., et al. (2009). Marginal mass islet transplantation with autologous mesenchymal stem cells promotes long-term islet allograft survival and sustained normoglycemia. Journal of Autoimmunity, 32, 116–124.PubMedCrossRefGoogle Scholar
  22. 22.
    Rosland, G. V., Svendsen, A., Torsvik, A., Sobala, E., McCormack, E., Immervoll, H., et al. (2009). Long-term cultures of bone marrow-derived human mesenchymal stem cells frequently undergo spontaneous malignant transformation. Cancer Research, 69, 5331–5339.PubMedCrossRefGoogle Scholar
  23. 23.
    Vacanti, V., Kong, E., Suzuki, G., Sato, K., Canty, J. M., & Lee, T. (2005). Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture. Journal of Cellular Physiology, 205, 194–201.PubMedCrossRefGoogle Scholar
  24. 24.
    Barry, F. P., & Murphy, J. M. (2004). Mesenchymal stem cells: clinical applications and biological characterization. The International Journal of Biochemistry & Cell Biology, 36, 568–584.CrossRefGoogle Scholar
  25. 25.
    Uccelli, A., Moretta, L., & Pistoia, V. (2008). Mesenchymal stem cells in health and disease. Nature Reviews Immunology, 8, 726–736.PubMedCrossRefGoogle Scholar
  26. 26.
    Jackson, K. A., Majka, S. M., Wang, H., Pocius, J., Hartley, C. J., Majesky, M. W., et al. (2001). Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells. The Journal of Clinical Investigation, 107, 1395–1402.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jiang Li
    • 1
    • 2
  • Mohamed B. Ezzelarab
    • 1
  • David Ayares
    • 3
  • David K. C. Cooper
    • 1
    Email author
  1. 1.The Thomas E. Starzl Transplantation InstituteUniversity of Pittsburgh Medical CenterPittsburghUSA
  2. 2.Department of Transplantation SurgeryTianjin First Central Hospital, Tianjin Medical UniversityTianjinChina
  3. 3.Revivicor Inc.BlacksburgUSA

Personalised recommendations