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Poster_NKT_Kit-Activation.pdf (656KB, MIME type: application/pdf)

Isolation and activation of human CD3+CD56+ NKT cells Ilka Bondzio, JÃ¼rgen Schmitz, Volker Huppert

Conclusion: A suitable small-scale, two-step method based on MACSÂ® Technology was developed for the separation of NKT cells from human PBMCs. In this process, monocytes and NK cells are first depleted according to their expression of CD14, NKp46, or CD16, after which NKT cells are enriched from the remaining cells by positive selection with CD56 MicroBeads.We showed that it is possible to achieve mean end-purities of 92 Â±5.9 %, though these values depend on the frequency NKT cells in the original, unseparated sample. Our data demonstrate that CD25 and/or CD69 expression of NKT cells isolated using MACSÂ® Technology can be induced with the use of a standard T cell activation protocol (T Cell Activation/Expansion Kit, human), which suggests that isolated NKT cells are functionally intact. In additional tests, we would like to evaluate whether it is also possible to expand separated NKT cells using a standard T cell expansion protocol that also uses MACSiBeadâ„¢ Particles.The commercially available NKT Cell Isolation Kit (Miltenyi Biotec) in conjunction with the T Cell Activation/ Expansion Kit may provide useful tools for NKT cell research, helping to better understand the roles of NK and NKT cells in the immune response of healthy donors and patients. An improved understanding may then lead to the development of novel treatment strategies, for example for different malignancies.

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(del) (cur) 01:30, 17 November 2006 . . Volker Huppert (671272 bytes) (Isolation and activation of human CD3+CD56+ NKT cells Ilka Bondzio, Jürgen Schmitz, Volker Huppert Conclusion: A suitable small-scale, two-step method based on MACS® Technology was developed for the separation of NKT cells from human PBMCs. In this pro)