European Heart Journal Advance Access originally published online on August 2, 2007
European Heart Journal 2007 28(17):2174-2175; doi:10.1093/eurheartj/ehm240
The REPAIR-AMI and ASTAMI trials: cell isolation procedures
Section of Transplantation Immunology
Institute of Immunology
Rikshospitalet Medical Center
Sognsvannsveien 20
Oslo
Norway
Ex Vivo Cell Laboratory
Institute of Immunology
Rikshospitalet Medical Center
Sogsnvannsveien 20
Oslo 0027
Norway
Tel: +47 2307 1379 Fax: +43 2307 3780 E-mail address: torstein.egeland{at}rikshospitalet.no
In a recent article, Seeger et al.1 compare the methods for the preparation of the bone marrow mononuclear cells (BM-MNC) in the REPAIR-AMI2 and ASTAMI3 trials.
In their paper, Seeger et al.1 claim to use Ficoll (Cambrex) for gradient centrifugation. Ficoll is a high-molecular-weight sucrose polymer, now rarely used on its own for cell separation procedures. Presumably, Seeger et al. mean Ficoll-Paque, which is Ficoll combined with sodium diatrizoate, density 1.077 g/mL. Ficoll-Paque or identical media from other producers is apparently the reagents used in their clinical studies, and they have identical constituents to the Lymphoprep density gradient medium used in the ASTAMI study.4,5
In the experiments described in Figures 1 and 2 and Table 1, Seeger et al.1 use practically identical experimental conditions to compare BM-MNC isolated by their ficoll solution and Lymphoprep. They find significant differences between the cell recoveries obtained using the two products, a difference which in turn is the only discernible reason for the differences observed for the number of CD45+/CD34+ BM-MNC, CFU, and MSC. Indeed, "... the overnight incubation protocols did not additionally affect the recovery of HSC, MSC and CFU (...), indicating that the reduced cell recovery between the two protocols is predominantly caused by differences in the initial centrifugation steps".1 As this is a central observation in their study, we believe that Seeger et al. should discuss how they were able to get different cell recoveries using the same bone marrow aspirates and identical gradient centrifugation media. Alternatively, if they actually used Ficoll, they need to explain why the density gradient medium used in this study was different from that used in their previous studies.1,2
References
- Seeger F, Tonn T, Krzossok N, Zeiher AM, Dimmeler S. Cell isolation procedures matter: a comparison of different isolation protocols of bone marrow mononuclear cells used for cell therapy in patients with acute myocaridial infarction. Eur Heart J (2007) 28:766–772.
[Abstract/Free Full Text] - Schächinger V, Erbs S, Elsässer A, Haberbosch W, Hambrecht R, Hölschermann H, Yu J, Corti R, Mathey DG, Hamm CW, Süselbeck T, Werner N, Haase J, Neuzner J, Germing A, Mark B, Assmus B, Tonn T, Dimmeler S, Zeiher AM, for the REPAIR-AMI trial. Improved clinical outcome after intracoronary administration of bone-marrow-derived progenitor cells in acute myocardial infarction: final 1-year results of the REPAIR-AMI trial. Eur Heart J (2006) 27:2775–2783.
[Abstract/Free Full Text] - Lunde K, Solheim S, Aakhus S, Arnesen H, Abdelnoor M, Egeland T, Endresen K, Ilebekk A, Mangschau A, Fjeld JG, Smith HJ, Taraldsrud E, Grøgaard HK, Bjørnerheim R, Brekke M, Müller C, Hopp E, Ragnarsson A, Brinchmann JE, Forfang K. Intracoronary injection of mononuclear bone marrow cells in acute myocardial infarction. N Engl J Med (2006) 355:1199–1209.
[Abstract/Free Full Text] - http://www5.gelifesciences.com/aptrix/upp01077.nsf/Content/Products?OpenDocument&parentid=17144002&moduleid=167164.
- http://www.axis-shield.com/densityhome/density/Data%20Sheet%20Lymphoprep.pdf.
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