Cell isolation and expansion using Dynabeads((R))
ABSTRACT This chapter describes the use of Dynabeads for cell isolation and expansion. Dynabeads are uniform polystyrene spherical beads that have been made magnetisable and superparamagnetic, meaning they are only magnetic in a magnetic field. Due to this property, the beads can easily be resuspended when the magnetic field is removed. The invention of Dynabeads made, by Professor John Ugelstad, has revolutionized the separation of many biological materials. For example, the attachment of target-specific antibodies to the surface of the beads allows capture and isolation of intact cells directly from a complex suspension such as blood. This is all accomplished under the influence of a simple magnetic field without the need for column separation techniques or centrifugation. In general, magnetic beads coated with specific antibodies can be used either for isolation or depletion of various cell types. Positive or negative cell isolation can be performed depending on the nature of the starting sample, the cell surface markers and the downstream application in question. Positive cell isolation is the method of choice for unprocessed samples, such as whole blood, and for downstream molecular applications. Positive cell isolation can also be used for any downstream application after detachment and removal of the beads. Negative cell isolation is the method of choice when it is critical that cells of interest remain untouched, i.e., no antibodies have been bound to any cell surface markers on the cells of interest. Some cell populations can only be defined by multiple cell surface markers. Such populations of cells can be isolated by the combination of negative and positive cell isolation. By coupling Dynabeads with antibodies directed against cell surface activation molecules, the beads can be used both for isolation and expansion of the cells. Dynabeads are currently used in two major clinical applications: 1) In the Isolex 300i Magnetic Cell Selection System for CD34 Stem Cell Isolation--2) For ex vivo T cell isolation and expansion using Dynabeads ClinExVivo CD3/CD28 for clinical trials in novel adoptive immunotherapy.
- SourceAvailable from: Martin BrandlBiomed 2012, Biomedical Engineering, Innsbruck, Austria; 02/2012
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ABSTRACT: Localized and metastatic cancers give rise to circulating tumor cells (CTCs) which are detectable in the bloodstream. Recent studies have highlighted the prognostic significance of the presence and number of CTCs, particularly in patients with metastatic disease. Future studies are designed not only to detect CTCs, but also to characterize them. This review discusses current and developing methodologies for the isolation and characterization of CTCs as well as recent studies focusing on the clinical relevance of CTC detection and characterization.American Journal of Cancer Research 01/2011; 1(6):740-51. · 3.97 Impact Factor
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ABSTRACT: Background/Aims: The aim of this study is to determine the relationship between circulating tumor cells (CTCs) and response to chemotherapy in patients with unresectable metastatic colorectal cancer. Methodology: CTCs of twelve consecutive patients with K-ras wild type colorectal cancer with synchronous and/or metachronous unresectable metastatic lesions were measured by the cell search system between January 2009 and December 2010. CTCs were measured before and after chemotherapy. The regimen consisted of four courses (three months) of SOX (TS-1+L-OHP) + panitumumab. Results: Four patients (33%) had no detectable CTCs before chemotherapy. For these patients, no CTCs were detected after chemotherapy and their serum carcinoembryonic antigen (CEA) levels decreased after chemotherapy. Four of the other eight patients with positive CTCs had no detectable CTCs after chemotherapy and their serum CEA levels decreased after chemotherapy. The other four of eight patients with positive CTCs continued to have CTCs after chemotherapy and all four patients died of cancer within eight months after starting chemotherapy. On the other hand, all eight patients without CTCs after chemotherapy were alive over a year after starting chemotherapy. Conclusions: CTCs may be able to predict the response to chemotherapy in patients with unresectable metastatic colorectal cancer.Hepato-gastroenterology 02/2012; 59(119). DOI:10.5754/hge11970 · 0.91 Impact Factor