- Short Communication
Homing of annexin-labeled stem cells to apoptotic cells
Cellular & Molecular Biology Letters volume 14, pages 100–112 (2009)
Ischemic diseases are characterized by the presence of pro-apoptotic stimuli, which initiate a cascade of processes that lead to cell injury and death. Several molecules and events represent detectable indicators of the different stages of apoptosis. Among these indicators is phosphatidylserine (PS) translocation from the inner to the outer leaflet of the plasma membrane, which can be detected by annexinV (ANXA5) conjugation. This is a widely used in vivo and in vitro assay marking the early stages of apoptosis. We report here on an original method that employs PS-ANXA5 conjugation to target stem cells to apoptotic cells. Mesenchymal stem cells (MSCs) from GFP-positive transgenic rats were biotinylated on membrane surfaces with sulfosuccinimidyl-6-(biotinamido) hexanoate (sulfo-NHS-LC-biot) and then bound to avidin. The avidin-biotinylated MSCs were labeled with biotin conjugated ANXA5. Bovine aortic endothelial cells (BAE-1 cells) were exposed to UVC to induce caspasedependent apoptosis. Finally, we tested the ability of ANXA5-labeled MSCs to bind BAE-1 apoptotic cells: suspended ANXA5-labeled MSCs were seeded for 1 hour on a monolayer of UV-treated or control BAE-1 cells. After washing, the number of MSCs bound to BAE-1 cells was evaluated by confocal microscopy. Statistical analysis demonstrated a significant increase in the number of MSCs tagged to apoptotic BAE-1 cells. Therefore, stem cell ANXA5 tagging via biotin-avidin bridges could be a straightforward method of improving homing to apoptotic tissues.
- BAE-1 cells:
bovine aortic endothelial cells
Dulbecco’s modified Eagle’s medium
fetal calf serum
green fluorescent protein
minimum essential Eagle’s medium, alpha modified
mesenchymal stem cells
phosphate buffer saline
Mimeault, M. and Batra, S.K. Concise review: recent advances on the significance of stem cells in tissue regeneration and cancer therapies. Stem Cells 24 (2006) 2319–2345.
Chen, C.P., Lee, Y.J., Chiu, S.T., Shyu, W.C., Lee, M.Y., Huang, S.P. and Li, H. The application of stem cells in the treatment of ischemic diseases. Histol. Histopathol. 21 (2006) 1209–1216.
Kucia, M., Reca, R., Miekus, K., Wanzeck, J., Wojakowski, W., Janowska-Wieczorek, A., Ratajczak, J. and Ratajczak, M.Z. Trafficking of normal stem cells and metastasis of cancer stem cells involve similar mechanisms: pivotal role of the SDF-1-CXCR4 axis. Stem Cells 23 (2005) 879–894.
Lopez Ponte, A., Marais, E., Gallay, N., Langonné, A., Delorme, B., Herault, O., Charbord, P. and Domenech, J. The in vitro migration capacity of human bone marrow mesenchymal stem cells: comparison of chemokine and growth factor chemotactic activities. Stem Cells 25 (2007) 1737–1745.
Liang Tang, Y., Qian, K., Zhang, Y.C., Shen, L. and Phillips, M.I. Mobilizing of haematopoietic stem cells to ischemic myocardium by plasmid mediated stromal-cell-derived factor-1α (SDF-1α) treatment. Regul. Pept. 125 (2005) 1–8.
Lum, L.G., Davol, P.A. and Lee, R.J. The new face of bispecific antibodies: targeting cancer and much more. Exp. Hematol. 34 (2006) 1–6.
Lopez-Neblina, F., Toledo, A.H. and Toledo-Pereyra, L.H. Molecular biology of apoptosis in ischemia and reperfusion. J. Invest. Surg. 18 (2005) 335–350.
Abbate, A., Bussani, R., Amin, M.S., Vetrovec, G.W. and Baldi A. Acute myocardial infarction and heart failure: role of apoptosis. Int. J. Biochem. Cell. Biol. 38 (2006) 1834–1840.
Huerta, S., Goulet, E.J., Huerta-Yepez, S. and Livingston, E.H. Screening and detection of apoptosis. J. Surg. Res. 139 (2007) 143–156.
Kietselaer, B.L., Reutelingsperger, C.P., Boersma, H.H., Heidendal, G.A., Liem, I.H., Crijns, H.J., Narula, J. and Hofstra, L. Noninvasive detection of programmed cell loss with 99mTc-labeled annexin A5 in heart failure. J. Nucl. Met. 48 (2007) 562–567.
Ito, T., Suzuki, A., Okabe, M., Imai, E. and Hori, M. Application of bone marrow-derived stem cells in experimental nephrology. Exp. Nephrol. 9 (2001) 444–450.
Javazon, E.H., Colter, D.C., Schwarz, E.J. and Prockop, D.J. Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells 19 (2001) 219–225.
Takasawa, R., Nakamura, H., Mori, T. and Tanuma, S. Differential apoptotic pathways in human keratinocyte HaCaT cells exposed to UVB and UVC. Apoptosis 10 (2005) 1121–1130.
Kenis, H., Hofstra, L. and Reutelingsperger, C.P. Annexin A5: shifting from a diagnostic towards a therapeutic realm. Cell. Mol. Life Sci. 64 (2007) 2859–62.
Boersma, H.H., Kietselaer, B.L., Stolk, L.M., Bennaghmouch, A., Hofstra, L., Narula, J., Heidendal, G.A. and Reutelingsperger, C.P. Past, present and future of annexin A5: from protein discovery to clinical applications. J. Nucl. Med. 46 (2005) 2035–2050.
Tanaka, K., Einaga, K., Tsuchiyama, H., Tait, J.F. and Fujikawa, K. Preparation and characterization of a disulfide-linked bioconjugate of annexin V with the B-chain of urokinase: an improved fibrinolytic agent targeted to phospholipids-containing thrombi. Biochemistry 35 (1996) 922–929.
Zimmermann, V.S., Bondanza, A., Rovere-Querini, P., Colombo, B., Sacchi, A., Fascio, U., Corti, A. and Manfredi, A.A. Characterisation of functional biotinylated TNF-alpha targeted to the membrane of apoptotic melanoma cells. J. Immunol. Methods 276 (2003) 79–87.
Magnani, M., Chiarantini, L. and Mancini, U. Preparation and characterization of biotinylated red blood cells. Biotechnol. Appl. Biochem. 20 (1994) 335–345.
Dominici, S., Laguardia, M.E., Serafini, G., Chiarantini, L., Fortini, C., Tripiciano, A., Brocca-Cofano, E., Scoglio, A., Caputo, A., Fiorelli, V., Gavioli, R., Cafaro, A., Ensoli, B. and Magnani, M. Red blood cell-mediated delivery of recombinant HIV-1 Tat protein in mice induces anti-Tat neutralizing antibodies and CTL. Vaccine 21 (2003) 2073–2081.
Suzuki, T. and Dale, G.L. Senescent erythrocytes: isolation of in vivo aged cells and their biochemical characteristic. Proc. Natl. Acad. Sci. USA 85 (1988) 1647–1651.
Hoya, K., Guterman, L.R., Miskolczi, L. and Hopkins, L.N. A novel intravascular drug delivery method using endothelial biotinylation and avidin-biotin binding. Drug Deliv. 8 (2001) 215–222.
A. Gerasimou, R. Ramella and A. Brero contributed equally to this paper.
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Gerasimou, A., Ramella, R., Brero, A. et al. Homing of annexin-labeled stem cells to apoptotic cells. Cell Mol Biol Lett 14, 100–112 (2009). https://doi.org/10.2478/s11658-008-0038-1
- Stem cell