Abstract
Magnetic beads (Dynabeads™) have been used for the purification of endothelial cells. One application for this procedure may be for single-stage seeding of bypass grafts. The number of endothelial cells (EC) isolated is crucial and therefore to increase the number of cells extracted, a higher number of Dynabeads™ per cell may need to be used. The effect of large numbers of CD31 Dynabeads™ on cell proliferation/metabolism is unknown. We undertook this study using CD31-coated Dynabeads™ and EC from human umbilical vein. EC were coated at concentrations of 4, 10, or 50 beads per cell. The cells were cultured for 6 days with control being normal EC. Cellular proliferation was assessed by trypsinization of cells and metabolism assessed with an Alamar blue™ viability assay. In a further experiment a compliant polyurethane graft was single-stage seeded with both coated Dynabeads™ and normal EC. The results showed that using a higher number of beads per cell resulted in a reduction in cell proliferation and a reduction in cell metabolism. The total number of Dynabeads™-coated cells in culture compared to controls (%) by day 6 were 30.7±2.56, 41.3±9.8 and 59.2±7.3 for 50, 10, and 4 beads per cell, respectively. The corresponding results for Alamar blue were 43.7±1.2, 61.8±1.4, and 72.1±4.3. The seeded grafts showed reduced metabolism with the Dynabeads™-coated EC. In conclusion, high numbers of beads per cell have a late detrimental effect on cell proliferation and metabolism. Therefore for single-stage seeding lower numbers of Dynabeads™ will need to be used with resultant reduction in the number of available EC.
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Arts CH, Hedeman Joosten PP, Blankensteijn JD, et al. Contaminants from the transplant contribute to intimal hyperplasia associated with microvascular endothelial cell seeding. Eur J Vasc Endovasc Surg. 2002;23:29–38.
Conrad-Lapostolle V, Bordenave L, Baquey C. Optimization of use of UEA-1 magnetic beads for endothelial cell isolation. Cell Biol Toxicol. 1996;12:189–97.
Deutsch M, Meinhart J, Fischlein T, Preiss P, Zilla P. Clinical autologous in vitro endothelialization of infrainguinal EPTFE grafts in 100 patients: a 9-year experience. Surgery. 1999;126:847–55.
Giudiceandrea A, Seifalian A M, Krijgsman B, Hamilton G. Effect of prolonged pulsatile shear stress in vitro on endothelial cell seeded PTFE and compliant polyurethane vascular grafts. Eur J Vasc Endovasc Surg. 1998;15:147–54.
Hewett PW, Murray JC. Immunomagnetic purification of human microvessel endothelial cells using Dynabeads coated with monoclonal antibodies to PECAM-1. Eur J Cell Biol. 1993;62:451–4.
Jackson CJ, Garbett PK, Nissen B, Schrieber L. Binding of human endothelium to Ulex europaeus I-coated Dynabeads: application to the isolation of microvascular endothelium. J Cell Sci. 1990;96 (Pt 2):257–62.
Laube HR, Duwe J, Rutsch W, Konertz W. Clinical experience with autologous endothelial cell-seeded polytetrafluoroethylene coronary artery bypass grafts. J Thorac Cardiovasc Surg. 2000;120:134–41.
Meerbaum, SO, Sharp WV, Schmidt SP. Lower extremity revascularization with polytetrafluoroethylene grafts seeded with microvascular endothelial cells. In: Zilla P, Fasol R, Callow A (eds). Applied cardiovascular biology. 1990:107–19.
Rosenman JE, Kempczinski RF, Pearce W H, Silberstein EB. Kinetics of endothelial cell seeding. J Vasc Surg. 1985;2:778–84.
Rubens FD, Labow RS, Meek E, Dudani AK, Ganz PR. Tissue factor expression by cells used for sodding of prosthetic vascular grafts. J Surg Res. 1997;72:22–8.
Salacinski HJ, Tai NR, Punshon G, Giudiceandrea A, Hamilton G, Seifalian AM. Optimal endothelialisation of a new compliant poly (varbonate-urea) urethane vascular graft with effect of physiological shear stress. Eur J Vasc Endovasc Surg. 2000;20:342–52.
Salacinski HJ, Tiwari A, Hamilton G, Seifalian AM. Cellular engineering of vascular bypass grafts: role of chemical coatings for enhancing endothelial cell attachment. Med Biol Eng Comput. 2001a;39:609–18.
Salacinski HJ, Tai NR, Carson RJ, Edwards A, Hamilton G, Seifalian AM. In vitro stability of a novel compliant poly (carbonate-urea) urethane to oxidative and hydrolytic stress. J Biomed Mater Res. 2001b;59:207–18.
Schmidt SP, Meerbaum SO, Anderson JM, Clarke RE, Zellers RA, Sharp WV. Evaluation of expanded polytetrafluoroethylene arteriovenous access grafts onto which microvessel-derived cells were transplanted to “improve” graft performance: preliminary results. Ann Vasc Surg. 1998;12:405–11.
Seifalian AM, Salacinski HJ, Punshon G, Krijgsman B, Hamilton G. A new technique for measuring the cell growth and metabolism of endothelial cells seeded on vascular prostheses. J Biomed Mater Res. 2001;55:637–44.
Seifalian AM, Tiwari A, Hamilton G, Salacinski HJ. Improving the Clinical patency of prosthetic vascular and coronary bypass grafts: the role of seeding and tissue engineering. Artif Organs. 2002;26:307–20.
Tiwari A, Salacinski HJ, Hamilton G, Seifalian AM. Tissue engineering of vascular bypass grafts: role of endothelial cell extraction. Eur J Vasc Endovasc Surg. 2001;21:193–201.
Tiwari A, Salacinski HJ, Punshon G, Hamilton G, Seifalian AM. Development of a hybrid cardiovascular graft using a tissue engineering approach. FASEB J. 2002;16:791–6.
Tiwari A, Cheng KS, Salacinski HJ, Hamilton G, Seifalian AM. Improving the patency of vascular bypass grafts: the role of suture materials and surgical techniques on reducing anastomotic compliance mismatch. Eur J Vasc Endovasc Surg. 2003a;25:287–95.
Tiwari A, Kidane A, Salacinski HJ, Punshon G, Hamilton G, Seifalian AM. Improving endothelial cell retention for single stage seeding of prosthetic grafts: use of polymer sequences of arginine-glycine-asparatate (RGD). Eur J Vasc Endovasc Surg. 2003b;25:325–9.
Vici M, Pasquinelli G, Preda P, et al. Electron microscopic and immunocytochemical profiles of human subcutaneous fat tissue microvascular endothelial cells. Ann Vasc Surg. 1993;7:541–8.
Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implication for cell based therapies. Tissue Eng. 2001;7:211–28.
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Tiwari, A., Punshon, G., Kidane, A. et al. Magnetic beads (Dynabead™) toxicity to endothelial cells at high bead concentration: Implication for tissue engineering of vascular prosthesis. Cell Biol Toxicol 19, 265–272 (2003). https://doi.org/10.1023/B:CBTO.0000004929.37511.ed
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DOI: https://doi.org/10.1023/B:CBTO.0000004929.37511.ed