Cell Biochemistry and Biophysics

, Volume 67, Issue 3, pp 1181–1191 | Cite as

Overexpression of the Mesenchymal Stem Cell Cxcr4 Gene in Irradiated Mice Increases the Homing Capacity of These Cells

  • Wei Chen
  • Miao Li
  • Hai Cheng
  • Zhiling Yan
  • Jiang Cao
  • Bin Pan
  • Wei Sang
  • Qingyun Wu
  • Lingyu Zeng
  • Zhenyu Li
  • Kailin XuEmail author
Original Paper


The efficiency of the intravascular delivery of mesenchymal stem cells (MSCs) homing to bone marrow has been largely limited. This study aimed to evaluate the homing efficacy in irradiated mice of MSCs that have been engineered to overexpress the murine Cxcr4 gene. Mouse MSCs were infected by a lentivirus vector carrying Cxcr4. MSC migration was detected by an in vitro transwell migration assay. EGFP-positive MSCs were systemically injected into BALB/c mice and detected in bone marrow samples by flow cytometry. The concentration of mouse stromal-derived factor 1 was detected by ELISA. The plasma concentration of the inflammatory cytokines, interleukin (IL)-6, IL-10, MCP-1, IFN-γ, TNF-α, and IL-12p70, were determined by cytometric bead array. MSCs that overexpressed Cxcr4 displayed enhanced migration toward a stromal-derived factor 1 gradient. The transplantation of Cxcr4-overexpressing MSCs into irradiated mice leads to increased homing to the bone marrow. Moreover, the frequency of the EGFP-positive cells in a bone marrow infusion 24 h after total body irradiation was 2.2-fold more than at 4 h after irradiation. The concentration of both plasma and bone marrow stromal-derived factor 1 increased after irradiation, and this was positively correlated with the number of Cxcr4-overexpressing MSCs homing to the bone marrow. Moreover, compared with the control groups, the plasma levels of IL-6, IFN-γ, TNF-α, and MCP-1 and IL-12p70 in recipients infused with Cxcr4-overexpressing MSCs was significantly decreased. The level of IL-10 was increased, which correlated with changes in the Th1 and Th2 subset balance. MSCs that overexpressed Cxcr4 and were injected into irradiated mice had an enhanced homing capacity which was related to the bone marrow level of stromal-derived factor 1.


Chemokine receptor 4 Lentivirus Mesenchymal stem cell Mice Stromal cell-derived factor 1 Total body irradiation 



The authors wish to acknowledge CuiPing Zhang and GuoLiang Song (Xuzhou medical College) for their technical assistance. The authors would like to thank Inder M. Verma (Salk Institute, San Diego, CA, USA) for kindly providing the LV-lacz and Shohei Hori for MIGR-1. This article was proofread by a native English professional with science background at Elixigen Corporation. This study was supported by Grants from the National Natural Science Foundation of China (No. 30971281 to K. Xu) and the Xuzhou Science and Technology Project, China (No. XZZD1138 to M. Li).


  1. 1.
    Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., Mosca, J. D., et al. (1999). Multilineage potential of adult human mesenchymal stem cells. Science, 284, 143–147.PubMedCrossRefGoogle Scholar
  2. 2.
    Parekkadan, B., & Milwid, J. M. (2010). Mesenchymal stem cells as therapeutics. Annual Review of Biomedical Engineering, 12, 87–117.PubMedCrossRefGoogle Scholar
  3. 3.
    English, K., French, A., & Wood, K. J. (2010). Mesenchymal stromal cells: Facilitators of successful transplantation? Cell Stem Cell, 7, 431–442.PubMedCrossRefGoogle Scholar
  4. 4.
    Wagner, J., Kean, T., Young, R., Dennis, J. E., & Caplan, A. I. (2009). Optimizing mesenchymal stem cell-based therapeutics. Current Opinion in Biotechnology, 20, 531–536.PubMedCrossRefGoogle Scholar
  5. 5.
    Barbash, I. M., Chouraqui, P., Baron, J., Feinberg, M. S., Etzion, S., Tessone, A., et al. (2003). Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium: Feasibility, cell migration, and body distribution. Circulation, 108, 863–868.PubMedCrossRefGoogle Scholar
  6. 6.
    Lee, R. H., Pulin, A. A., Seo, M. J., Kota, D. J., Ylostalo, J., Larson, B. L., et al. (2009). Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6. Cell Stem Cell, 5, 54–63.PubMedCrossRefGoogle Scholar
  7. 7.
    Gao, J., Dennis, J. E., Muzic, R. F., Lundberg, M., & Caplan, A. I. (2001). The dynamic in vivo distribution of bone marrow-derived mesenchymal stem cells after infusion. Cells Tissues Organs, 169, 12–20.PubMedCrossRefGoogle Scholar
  8. 8.
    Karp, J. M., & Leng Teo, G. S. (2009). Mesenchymal stem cell homing: The devil is in the details. Cell Stem Cell, 4, 206–216.PubMedCrossRefGoogle Scholar
  9. 9.
    Nombela-Arrieta, C., Ritz, J., & Silberstein, L. E. (2011). The elusive nature and function of mesenchymal stem cells. Nature Reviews Molecular Cell Biology, 12, 126–131.PubMedCrossRefGoogle Scholar
  10. 10.
    Lapidot, T., Dar, A., & Kollet, O. (2005). How do stem cells find their way home? Blood, 106, 1901–1910.PubMedCrossRefGoogle Scholar
  11. 11.
    Wynn, R. F., Hart, C. A., Corradi-Perini, C., O’Neill, L., Evans, C. A., Wraith, J. E., et al. (2004). A small proportion of mesenchymal stem cells strongly expresses functionally active CXCR4 receptor capable of promoting migration to bone marrow. Blood, 104, 2643–2645.PubMedCrossRefGoogle Scholar
  12. 12.
    Honczarenko, M., Le, Y., Swierkowski, M., Ghiran, I., Glodek, A. M., & Silberstein, L. E. (2006). Human bone marrow stromal cells express a distinct set of biologically functional chemokine receptors. Stem Cells, 24, 1030–1041.PubMedCrossRefGoogle Scholar
  13. 13.
    Son, B. R., Marquez-Curtis, L. A., Kucia, M., Wysoczynski, M., Turner, A. R., Ratajczak, J., et al. (2006). Migration of bone marrow and cord blood mesenchymal stem cells in vitro is regulated by stromal-derived factor-1-CXCR4 and hepatocyte growth factor-c-met axes and involves matrix metalloproteinases. Stem Cells, 24, 1254–1264.PubMedCrossRefGoogle Scholar
  14. 14.
    Ponte, A. L., Marais, E., Gallay, N., Langonne, A., Delorme, B., Herault, O., et al. (2007). The in vitro migration capacity of human bone marrow mesenchymal stem cells: Comparison of chemokine and growth factor chemotactic activities. Stem Cells, 25, 1737–1745.PubMedCrossRefGoogle Scholar
  15. 15.
    Rombouts, W. J., & Ploemacher, R. E. (2003). Primary murine MSC show highly efficient homing to the bone marrow but lose homing ability following culture. Leukemia, 17, 160–170.PubMedCrossRefGoogle Scholar
  16. 16.
    Mazzinghi, B., Ronconi, E., Lazzeri, E., Sagrinati, C., Ballerini, L., Angelotti, M. L., et al. (2008). Essential but differential role for CXCR4 and CXCR7 in the therapeutic homing of human renal progenitor cells. Journal of Experimental Medicine, 205, 479–490.PubMedCrossRefGoogle Scholar
  17. 17.
    Kyriakou, C., Rabin, N., Pizzey, A., Nathwani, A., & Yong, K. (2008). Factors that influence short-term homing of human bone marrow-derived mesenchymal stem cells in a xenogeneic animal model. Haematologica, 93, 1457–1465.PubMedCrossRefGoogle Scholar
  18. 18.
    Cheng, Z., Ou, L., Zhou, X., Li, F., Jia, X., Zhang, Y., et al. (2008). Targeted migration of mesenchymal stem cells modified with CXCR4 gene to infarcted myocardium improves cardiac performance. Molecular Therapy, 16, 571–579.PubMedCrossRefGoogle Scholar
  19. 19.
    Cho, S. W., Sun, H. J., Yang, J. Y., Jung, J. Y., An, J. H., Cho, H. Y., et al. (2009). Transplantation of mesenchymal stem cells overexpressing RANK-Fc or CXCR4 prevents bone loss in ovariectomized mice. Molecular Therapy, 17, 1979–1987.PubMedCrossRefGoogle Scholar
  20. 20.
    Ricks, D. M., Kutner, R., Zhang, X. Y., Welsh, D. A., & Reiser, J. (2008). Optimized lentiviral transduction of mouse bone marrow-derived mesenchymal stem cells. Stem Cells Dev, 17, 441–450.PubMedCrossRefGoogle Scholar
  21. 21.
    Bobis-Wozowicz, S., Miekus, K., Wybieralska, E., Jarocha, D., Zawisz, A., Madeja, Z., et al. (2011). Genetically modified adipose tissue-derived mesenchymal stem cells overexpressing CXCR4 display increased motility, invasiveness, and homing to bone marrow of NOD/SCID mice. Experimental Hematology, 39(686–696), e684.Google Scholar
  22. 22.
    Zhang, D., Fan, G. C., Zhou, X., Zhao, T., Pasha, Z., Xu, M., et al. (2008). Over-expression of CXCR4 on mesenchymal stem cells augments myoangiogenesis in the infarcted myocardium. Journal of Molecular and Cellular Cardiology, 44, 281–292.PubMedCrossRefGoogle Scholar
  23. 23.
    Dar, A., Goichberg, P., Shinder, V., Kalinkovich, A., Kollet, O., Netzer, N., et al. (2005). Chemokine receptor CXCR4-dependent internalization and resecretion of functional chemokine SDF-1 by bone marrow endothelial and stromal cells. Nature Immunology, 6, 1038–1046.PubMedCrossRefGoogle Scholar
  24. 24.
    Kollet, O., Shivtiel, S., Chen, Y. Q., Suriawinata, J., Thung, S. N., Dabeva, M. D., et al. (2003). HGF, SDF-1, and MMP-9 are involved in stress-induced human CD34+ stem cell recruitment to the liver. Journal of Clinical Investigation, 112, 160–169.PubMedGoogle Scholar
  25. 25.
    Shi, M., Li, J., Liao, L., Chen, B., Li, B., Chen, L., et al. (2007). Regulation of CXCR4 expression in human mesenchymal stem cells by cytokine treatment: Role in homing efficiency in NOD/SCID mice. Haematologica, 92, 897–904.PubMedCrossRefGoogle Scholar
  26. 26.
    Liu, H., Liu, S., Li, Y., Wang, X., Xue, W., Ge, G., et al. (2012). The role of SDF-1-CXCR4/CXCR7 axis in the therapeutic effects of hypoxia-preconditioned mesenchymal stem cells for renal ischemia/reperfusion injury. PLoS One, 7, e34608.PubMedCrossRefGoogle Scholar
  27. 27.
    Gheisari, Y., Azadmanesh, K., Ahmadbeigi, N., Nassiri, S. M., Golestaneh, A. F., Naderi, M., et al. (2012). Genetic modification of mesenchymal stem cells to overexpress CXCR4 and CXCR7 does not improve the homing and therapeutic potentials of these cells in experimental acute kidney injury. Stem Cells Dev, 21(6), 2969–2980.PubMedCrossRefGoogle Scholar
  28. 28.
    Xun, C. Q., Thompson, J. S., Jennings, C. D., Brown, S. A., & Widmer, M. B. (1994). Effect of total body irradiation, busulfan-cyclophosphamide, or cyclophosphamide conditioning on inflammatory cytokine release and development of acute and chronic graft-versus-host disease in H-2-incompatible transplanted SCID mice. Blood, 83, 2360–2367.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Wei Chen
    • 1
  • Miao Li
    • 2
  • Hai Cheng
    • 1
  • Zhiling Yan
    • 1
  • Jiang Cao
    • 1
  • Bin Pan
    • 1
  • Wei Sang
    • 1
  • Qingyun Wu
    • 1
  • Lingyu Zeng
    • 1
  • Zhenyu Li
    • 1
  • Kailin Xu
    • 1
    Email author
  1. 1.Department of HematologyThe Affiliated Hospital of Xuzhou Medical CollegeXuzhouPeople’s Republic of China
  2. 2.Xuzhou Children’s HospitalXuzhouPeople’s Republic of China

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