by Andrea Armstead AL
A Novel Hydrogel-Based Approach for Cell Isolation, Purification and Release
The purification and enrichment of specific cell populations from complex starting samples is a critical component of multiple life science workflows. While the key objective of most protocols is to obtain a highly pure and viable population of cells for downstream applications and analysis, most do not meet this goal.
Issues with conventional cell separation methods, such as traditional mag- netic beads and fluorescent activated cell sorting, include low recovery and poor cell viability, high costs and lengthy protocol times. Magnetic beads can remain associated with the isolated cells, causing decreased cell viability and changes to cellular function, and may interfere with downstream post-processing assays. In the quest to deliver new cell- based therapeutics, it is necessary to be able to efficiently scale up cell separation workflows to process higher volumes of samples that meet emerging regulatory requirements.
Addressing the limitations of traditional cell
separation techniques QuickGel hydrogel (Quad Technologies, Woburn, Mass.) allows complete cell release from magnetic carrier particles at the end of the separation process. The addition of a biologically friendly buffer facilitates the in- stantaneous release of target cells from magnetic carriers. This leaves captured cells behind in their native state after removal of magnetic
Figure 1 – Comparison of conventional magnetic cell separation methods and the MagCloudz approach.
AMERICAN LABORATORY 40
Figure 2 – Representative flow cytometry dot plots for T-cell enrichment and purification from peripheral blood. a) Forward and side-scatter pro- file of the initial PBMC population; b) 56% CD3+/CD45+ T-cells (target cell population) were present in the initial sample; c) 92% T-cell uptake as indicated by the low percentage of T-cells remaining in the unbound cell fraction following MagCloudz streptavidin binding; d) forward and side- scatter profile of the recovered CD3+ T-cell population; e) high-purity T-cells were recovered from the MagCloudz streptavidin kit, enriched from 56% in the initial population to 98% in the final isolated/released population; and f) high viability (99%) of the recovered T-cells, indicated by the absence of annexin-V and propidium iodide staining in the bottom left quadrant of the plot. Eighty percent of the target T-cell population was recovered as magnetic label-free T-cells from the MagCloudz strep- tavidin kit in this assay.
MARCH 2016
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