Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8418090 | Journal of Immunological Methods | 2013 | 9 Pages |
Abstract
The TACs/Sep method significantly decreased the time required for NK cell isolation (1Â h vs. 4Â h), but resulted in higher red blood cell contamination. NK cell activation marker expression (including CD94, NKG2D, NKp30, NKp46, DNAM-1, 2B4, KIR2DL1/S1, KIR2DL2/L3, intracellular granzyme B, and perforin) was similar when comparing NK cells isolated by the TACs/Sep or DG/MACS methods, but the TACs/Sep method induced higher expression of CD16. In vitro cytotoxicity against HT29 colon cancer and K562 leukemia cells was not affected by the isolation method. Lastly, by combining the TACs/Sep NK cell isolation method with calcein-acetoxymethyl diacetylester (calcein-AM) release, the time required to assess in vitro cytotoxicity was reduced by 33% (4Â h) compared to protocols employing DG/MACS and chromium release. Altogether, these results provide the foundation for the development of a rapid, high throughput functional assay, and make it practical for the multiplexing of downstream applications, such as flow cytometric analysis and enzyme-linked immunosorbent assays (ELISAs).
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Authors
Edward C. So, Michelle A. Sallin, Xiaoyu Zhang, Siaw L. Chan, Lepakshi Sahni, Dan H. Schulze, Eduardo Davila, Scott E. Strome, Ajay Jain,