Image:Poster ISCT 06 Ilka print.pdf

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Poster_ISCT_06_Ilka_print.pdf (344KB, MIME type: application/pdf)

Development of an isolation method for CD3+/CD56+ NKT cells I. Bondzio, A. Arendt, J. Schmitz, V. Huppert

Conclusion A suitable small-scale two-step method based on MACS® Technology was developed for the separation of NKT cells from a PBMC sample. In this process, monocytes and NK cells are first depleted according to their expression of CD14, NKp46 and /or CD16. Then the remaining NK T cells are enriched by a positive selection with CD56 MicroBeads. It was demonstrated that it is possible to achieve final purities of more than 90%, depending on the NKT cell frequency in the unseparated sample. Additionally, the results of this new method are comparable to results obtained with the already available MACS CD56 MultiSort Kit. In the next development step the method has to be evaluated with many different donor samples for method validation. Furthermore, we will demonstrate that the autoMACS Separator (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) can be used for this NKT cell separation procedure. A commercially available “NK T Cell Isolation Kit ” (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) may provide a useful tool for NKT cell research. Data on NKT cell func tion achieved by using this method may provide arguments for either maintaining the CD3–/CD56+ NK cell isolation procedure or simplifying to NK/NK T isolation by CD56 enrichment only for clinical-scale NK cell preparation.

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  • (del) (cur) 01:27, 17 November 2006 . . Volker Huppert (352079 bytes) (Development of an isolation method for CD3+/CD56+ NKT cells I. Bondzio, A. Arendt, J. Schmitz, V. Huppert Conclusion A suitable small-scale two-step method based on MACS® Technology was developed for the separation of NKT cells from a PBMC sample. In th)

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